WO2024078118A1

WO2024078118A1 - Chemical reprogramming and pluripotent stem cells

Info

Publication number: WO2024078118A1
Application number: PCT/CN2023/113080
Authority: WO
Inventors: Hongkui Deng; Zhengyuan ZHANG; Huanjing HE; Yanglu WANG; Xinxing ZHONG; Guan Wang; Jinlin Wang; Jingyang GUAN
Original assignee: Peking University
Priority date: 2022-10-12
Filing date: 2023-08-15
Publication date: 2024-04-18

Abstract

Provided herein, in some aspects, are methods and compositions for cell conversion. The methods may convert a cell population comprising one cell type to another cell population comprising another cell type. The converted cell types may have increased cell differentiation potential. The converted cell types can comprise pluripotent stem cells. The compositions provided herein may comprise chemical reprogramming factors for converting cells. The compositions provided herein may comprise chemical reprogramming factors and cells. Also provided herein are reagents for carrying out the methods for converting cells. Additionally, provided herein are methods and compositions for using various cell types obtained by the methods and/or compositions provided herein.

Description

CHEMICAL REPROGRAMMING AND PLURIPOTENT STEM CELLS

BACKGROUND

There remains a significant need for cell resources for applications in basic research, therapeutics, agriculture, and food industry. Improved methods for industrial scale production of stem cells, as well as cells of various differentiation states and cell types, remain to be developed.

Cell identity can be established during development to acquire and maintain specialized cellular functions in somatic cells. Cellular reprogramming can manipulate cell identity, thereby enabling the generation of desired cell types that provide broad applications in disease modelling, drug discovery and regenerative medicine. Using cellular factors, including oocyte components and transcription factors, mouse and human somatic cells can be reprogrammed into pluripotent stem cells. Alternatively, chemical reprogramming can be utilized to induce somatic cells into pluripotent stem cells by simple exposure to small molecules. However, efficiency and kinetics of human chemical reprogramming system needs to be improved to robustly induce pluripotent stem cells from human somatic cells.

SUMMARY

Provided herein, in some aspects, are methods and compositions for cell conversion using chemical reprogramming factors. The methods and compositions may generate cells with enhanced differentiation potentials. The methods and compositions may convert a cell population comprising at least a cell type to another cell population comprising another cell type that exhibits increased differentiation potentials. Various cells generated by the methods and/or compositions may comprise pluripotent stem cells or other intermediate cells with increased potential to become pluripotent stem cells relative to the cell that is not converted. These intermediate cells may comprise intermediate plastic state cells, progenies thereof, or derivatives thereof.

The methods and compositions may bypass using genetic modification to generate cells with enhanced differentiation potentials. In some cases, not utilizing genetic modification to generate stem cells or cells of various differentiation states may decrease the negative impact of the genetic modification. Such negative impact may comprise accidental induction of mutations in the stem cells. In some cases, such negative impacts may also be generated by exogenous nucleic acid molecules or sequences used in the genetic modification of the cells. Cells comprising the accidental mutation (s) may have various undesirable properties, including but limited to, enhanced or unregulated cell proliferation potentials (that can lead to neoplastic diseases such as cancers) , unforeseen differentiation properties, and/or undesirable cell senescence stages. Utilizing chemical reprogramming factors, the methods and compositions provided herein can eliminate these negative impacts induced by the genetic modification. Furthermore, wherein when genetic modification needs to be utilized in downstream purposes after generation of converted cells, the methods and compositions provided herein can reduce the negative impacts described herein.

Using the chemical programming factors described herein, the methods and compositions can also increase the scalability for producing stem cells as well as cells of various differentiation states. These methods and compositions can generate stem cells or cells of various differentiation states within a shorter period of time, compared to existing methods. Hence, the methods and compositions can satisfy the need for cell resources for applications in basic research, therapeutics, agriculture, and food industry.

Provided herein, in some aspects, is a method for producing pluripotent stem cells. In some cases, method for producing pluripotent stem cells comprises: (a) contacting somatic cells with a first composition comprising one or more of (1) a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor; (2) a CBP/p300 bromodomain inhibitor; (3) a Menin-MLL interaction inhibitor; (4) a serine-threonine kinase Akt inhibitor; (5) a glycogen kinase inhibitor; (6) a TGFβ receptor inhibitor; (7) a retinoic acid receptor agonist; (8) an agonist for the G protein-coupled receptor Smoothened; (9) a Dot1L inhibitor; (10) a Jak1/Jak2 inhibitor; (11) an SAH hydrolase inhibitor; or (12) a c-Jun kinase inhibitor, thereby obtaining a first cell population; and (b) contacting the first cell population with a second composition comprising one or more of (i) a MEK inhibitor; (ii) a B-Raf inhibitor; or (iii) a histone deacetylase inhibitor, thereby obtaining a second cell population comprising pluripotent stem cells.

In some cases, the first composition comprises one or more of the c-Jun kinase inhibitor, the CBP/p300 bromodomain inhibitor, or the serine-threonine kinase Akt inhibitor. In some cases, the first composition comprises the c-Jun kinase inhibitor, the CBP/p300 bromodomain inhibitor, and the serine-threonine kinase Akt inhibitor. In some cases, the first composition comprises one or more of: the ROCK inhibitor; the CBP/p300 bromodomain inhibitor; the Menin-MLL interaction inhibitor; or the serine-threonine kinase Akt inhibitor. In some cases, the first composition comprises one or more of: the ROCK inhibitor, the c-Jun kinase inhibitor, or the CBP/p300 bromodomain inhibitor. In some cases, the first composition comprises (1) the Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor; (2) the CBP/p300 bromodomain inhibitor; (3) the Menin-MLL interaction inhibitor; (4) the serine-threonine kinase Akt inhibitor; (5) the glycogen kinase inhibitor; (6) the TGFβ receptor inhibitor; (7) the retinoic acid receptor agonist; (8) an agonist for the G protein-coupled receptor Smoothened; (9) the Dot1L inhibitor; (10) the Jak1/Jak2 inhibitor; (11) the SAH hydrolase inhibitor; and (12) the c-Jun kinase inhibitor. In some cases, the first composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor. In some cases, the first composition further comprises a G9a inhibitor or a DNMT inhibitor. In some cases, the first composition further comprises an adeno-sine kinase inhibitor, a casein kinase 2 inhibitor, or both.

In some cases, the second composition further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor. In some cases, the second composition further comprises one or more of an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor. In some cases, the second composition further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, or a ROCK inhibitor.

In some cases, the first cell population comprises intermediate plastic state cells that express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2. In some cases, the intermediate plastic state cells express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1. In some cases, the intermediate plastic state cells further express one or more of FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

In some cases, the pluripotent stem cells express one or more of OCT4, SOX2, or NANOG. In some cases, the pluripotent stem cells further express one or more of FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DPPA5, DNMT3L, REX1, or UTF1. In some cases, the pluripotent stem cells further express one or more of FGF4, ZFP57, DPPA5, or REX1. In some cases, the pluripotent stem cells further express one or more of DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DPPA5, DNMT3L, REX1, or UTF1.

In some cases, the somatic cells comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) . In some cases, the somatic cells comprise fibroblasts.

In some cases, the method converts the somatic cells into the pluripotent stem cells within less about 50 days. In some cases, the method converts the somatic cells into the pluripotent stem cells within at most about 32 days. In some cases, the method converts the somatic cells into the pluripotent stem cells within at most about 24 days. In some cases, the method converts the somatic cells into the pluripotent stem cells within at most about 14 days. In some cases, the somatic cells comprise a genetic modification. In some cases, the pluripotent stem cells comprise the genetic modification. In some cases, the genetic modification comprises an exogenous nucleic acid sequence. In some cases, the exogenous nucleic acid sequence encodes a polypeptide. In some cases, the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule. In some cases, the genetic modification comprises alteration of a genomic sequence. In some cases, the genetic modification reduces immunogenicity of the pluripotent stem cells.

In some cases, the ROCK inhibitor comprises Y-27632 or thiazovivin. In some cases, the ROCK inhibitor comprises Y-27632. In some cases, Y-27632 is present at about 1 μM to about 100 μM within the first composition or the second composition. In some cases, Y-27632 is present at about 2 μM to about 50 μM within the first composition or the second composition. In some cases, Y-27632 is present at about 4 μM to about 25 μM within the first composition or the second composition. In some cases, Y-27632 is present at about 10 μM within the first composition or the second composition. In some cases, the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409. In some cases, the CBP/p300 bromodomain inhibitor comprises SGC-CBP30. In some cases, SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM within the first composition. In some cases, SGC-CBP30 is present at about 0.4 μM to about 10 μM within the first composition. In some cases, SGC-CBP30 is present at about 0.8 μM to about 5 μM within the first composition. In some cases, SGC-CBP30 is present at about 2 μM within the first composition. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469. In some cases, VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM within the first composition. In some cases, VTP50469 is present at about 0.1 μM to about 2.5 μM within the first composition. In some cases, VTP50469 is present at about 0.2 μM to about 1.25 μM within the first composition. In some cases, VTP50469 is present at about 0.5 μM within the first composition. In some cases, the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor. In some cases, AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM within the first composition. In some cases, AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM within the first composition. In some cases, AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM within the first composition. In some cases, AKT Kinase Inhibitor is present at about 1 μM within the first composition. In some cases, the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014. In some cases, the glycogen kinase inhibitor comprises CHIR99021. In some cases, CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM within the first composition or the second composition. In some cases, CHIR99021 is present at about 1 μM to about 25 μM within the first composition or the second composition. In some cases, CHIR99021 is present at about 2 μM to about 12.5 μM within the first composition. In some cases, CHIR99021 is present at about 5 μM within the first composition. In some cases, CHIR99021 is present at about 0.75 μM to about 5 μM within the second composition. In some cases, CHIR99021 is present at about 1 μM to about 3 μM within the second composition. In some cases, CHIR99021 is present at about 2 μM within the second composition. In some cases, the TGFβ receptor inhibitor is an ALK5 inhibitor. In some cases, the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334, In some cases, the TGFβ receptor inhibitor comprises E-616452. In some cases, E-616452 is present at about 1 micromolar (μM) to about 100 μM within the first composition. In some cases, E-616452 is present at about 2 μM to about 50 μM within the first composition. In some cases, E-616452 is present at about 4 μM to about 25 μM within the first composition. In some cases, E-616452 is present at about 10 μM within the first composition. In some cases, the RAR agonist comprises TTNPB, Ch55, or AM580. In some cases, the RAR agonist comprises TTNPB. In some cases, TTNPB is present at about 0.2 μM to about 20 μM within the first composition. In some cases, TTNPB is present at about 0.4 μM to about 10 μM within the first composition. In some cases, TTNPB is present at about 0.8 μM to about 5 μM within the first composition. In some cases, TTNPB is present at about 2 μM within the first composition. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG. In some cases, SAG is present at about 0.05 micromolar (μM) to about 5 μM within the first composition. In some cases, SAG is present at about 0.1 μM to about 2.5 μM within the first composition In some cases, SAG is present at about 0.2 μM to about 1.25 μM within the first composition. In some cases, SAG is present at about 0.5 μM within the first composition. In some cases, the Dot1L inhibitor comprises EPZ004777 or EPZ5676. In some cases, the Dot1L inhibitor comprises the EPZ5676. In some cases, EPZ5676 is present at about 0.2 μM to about 20 μM within the first composition or the second composition. In some cases, EPZ5676 is present at about 0.4 μM to about 10 μM within the first composition or the second composition. In some cases, EPZ5676 is present at about 0.8 μM to about 5 μM within the first composition or the second composition. In some cases, EPZ5676 is present at about 2 μM within the first composition or the second composition. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib. In some cases, Ruxolitinib is present at about 0.1 μM to about 10 μM within the first composition. In some cases, Ruxolitinib is present at about 0.2 μM to about 5 μM within the first composition. In some cases, Ruxolitinib is present at about 0.4 μM to about 2.5 μM within the first composition. In some cases, Ruxolitinib is present at about 1 μM within the first composition. In some cases, the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA. In some cases, the SAH hydrolase inhibitor comprises DZNep. In some cases, DZNep is present at about 0.02 μM to about 2 μM within the first composition or the second composition. In some cases, DZNep is present at about 0.04 μM to about 1 μM within the first composition or the second composition. In some cases, DZNep is present at about 0.08 μM to about 0.5 μM within the first composition or the second composition. In some cases, DZNep is present at about 0.2 μM with the first composition or the second composition. In some cases, the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12. In some cases, the c-Jun kinase inhibitor comprises JNKIN8. In some cases, JNKIN8 is present at about 0.05 micromolar (μM) to about 5 μM within the first composition. In some cases, JNKIN8 is present at about 0.1 μM to about 2.5 μM within the first composition. In some cases, JNKIN8 is present at about 0.2 μM to about 1.25 μM within the first composition. In some cases, JNKIN8 is present at about 0.5 μM within the first composition. In some cases, the MEK inhibitor comprises PD0325901, AZD8330, or TAK-733. In some cases, the MEK inhibitor comprises PD0325901. In some cases, PD0325901 is present at about 0.1 μM to about 10 μM with the second composition. In some cases, PD0325901 is present at about 0.2 μM to about 5 μM with the second composition. In some cases, PD0325901 is present at about 0.4 μM to about 2.5 μM with the second composition. In some cases, PD0325901 is present at about 1 μM with the second composition. In some cases, the B-Raf inhibitor comprises SB590885, Vemurafenib, RAF265, or PLX4720. In some cases, the B-Raf inhibitor comprises SB590885. In some cases, SB590885 is present at about 0.05 μM to about 5 μM with the second composition. In some cases, SB590885 is present at about 0.1 μM to about 2.5μM with the second composition. In some cases, SB590885 is present at about 0.2 μM to about 1.25 μM with the second composition. In some cases, SB590885 is present at about 0.5 μM with the second composition. In some cases, the histone deacetylase inhibitor comprises valproic acid (VPA) , LMK235, MS275, or HDACi IV. In some cases, the histone deacetylase inhibitor comprises valproic acid (VPA) . In some cases, VPA is present at about 0.1 millimolar (mM) to about 10 mM within the second composition. In some cases, VPA is present at about 0.2 mM to about 5 mM within the second composition. In some cases, VPA is present at about 0.4 mM to about 2.5 mM within the second composition. In some cases, VPA is present at about 1 mM within the second composition. In some cases, the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702. In some cases, the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) . In some cases, 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM within the first composition. In some cases, 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM within the first composition. In some cases, 5-ITU is present at about 0.2 micromolar μM to about 1 μM within the first composition. In some cases, the 5-ITU is present at about 0.5 μM within the first composition. In some cases, the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid. In some cases, the casein kinase 2 inhibitor comprises the CX-4945. In some cases, CX-4945 is present at about 0.1 μM about 10 μM within the first composition. In some cases, CX-4945 is present at about 0.2 μM about 5 μM within the first composition. In some cases, CX-4945 is present at about 0.5 μM about 1 μM within the first composition. In some cases, the G9a inhibitor comprises Unc0224, Unc0638, Unc0642, or Bix01294. In some cases, the G9a inhibitor comprises Unc0224. In some cases, Unc0224 is present at about 0.1 μM to about 10 μM within the first composition. In some cases, Unc0224 is present at about 0.2 μM to about 5 μM within the first composition. In some cases, Unc0224 is present at about 0.5 μM to about 2 μM within the first composition. In some cases, Unc0224 is present at about 1 μM within the first composition. In some cases, the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027. In some cases, the DNMT inhibitor comprises 5-Azacytidine. In some cases, 5-Azacytidine is present at about 0.2 μM to about 20 μM within the first composition. In some cases, 5-Azacytidine is present at about 0.5 μM to about 10 μM within the first composition. In some cases, 5-Azacytidine is present at about 1 μM to about 5 μM within the first composition. In some cases, 5-Azacytidine is present at about 2 μM within the first composition. In some cases, the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190. In some cases, the p38 MAPK inhibitor comprises BIRB796. In some cases, BIRB796 is present at about 0.2 μM to about 20 μM within the first composition. In some cases, BIRB796 is present at about 0.4 μM to about 10 μM within the first composition. In some cases, BIRB796 is present at about 0.8 μM to about 5 μM within the first composition. In some cases, BIRB796 is present at about 2 μM within the first composition. In some cases, the inhibitor of histone demethylation comprises Tranylcypromine. In some cases, Tranylcypromine is present at about 1 μM to about 100 μM within the second composition. In some cases, Tranylcypromine is present at about 2 μM to about 50 μM within the second composition. In some cases, Tranylcypromine is present at about 4 μM to about 25 μM within the second composition. In some cases, Tranylcypromine is present at about 10 μM within the second composition. In some cases, the Wnt inhibitor comprises IWR-1 or IWP-2. In some cases, the Wnt inhibitor comprises IWR-1. In some cases, the Wnt inhibitor comprises IWP-2. In some cases, IWP-2 is present at about 0.2 μM to about 20 μM within the second composition. In some cases, IWP-2 is present at about 0.4 μM to about 10 μM within the second composition. In some cases, IWP-2 is present at about 0.8 μM to about 5 μM within the second composition. In some cases, IWP-2 is present at about 2 μM within the second composition.

Provided herein, in some aspects, are methods for reprogramming somatic cells. In some aspects, a method for reprogramming somatic cells comprises contacting the somatic cells with a composition comprising a c-Jun kinase inhibitor or a CBP/p300 bromodomain inhibitor. In some cases, the composition further comprises a serine-threonine kinase Akt inhibitor.

Provided herein, in some aspects, are methods for reprogramming somatic cells, comprising contacting the somatic cells with a composition comprising one or more of a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor.

In some cases, the composition further comprises a ROCK inhibitor or a Menin-MLL interaction inhibitor. In some cases, the composition further comprises one or more of: (1) a glycogen kinase inhibitor; (2) a TGFβ receptor inhibitor; (3) a retinoic acid receptor agonist; (4) an agonist for the G protein-coupled receptor Smoothened; (5) a Dot1L inhibitor; (6) a Jak1/Jak2 inhibitor; or (7) an SAH hydrolase inhibitor. In some cases, wherein the composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, a p38 MAPK inhibitor. In some cases, the composition further comprises a G9a inhibitor or a DNMT inhibitor. In some cases, the composition comprises the adenosine kinase inhibitor, the casein kinase 2 inhibitor, or both.

In some cases, the method induces conversion of the somatic cells into intermediate plastic state cells that express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2. In some cases, the intermediate plastic state cells express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1. In some cases, the intermediate plastic state cells further express one or more of FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

In some cases, the somatic cells comprise a genetic modification. In some cases, the intermediate plastic state cells comprise the genetic modification. In some cases, the genetic modification comprises an exogenous nucleic acid sequence. In some cases, the exogenous nucleic acid sequence encodes a polypeptide. In some cases, the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule. In some cases, the genetic modification comprises alteration of a genomic sequence. In some cases, the genetic modification reduces immunogenicity of the intermediate plastic state cells.

In some cases, the ROCK inhibitor comprises Y-27632 or thiazovivin. In some cases, the ROCK inhibitor comprises Y-27632. In some cases, Y-27632 is present at about 1 μM to about 100 μM within the composition. In some cases, Y-27632 is present at about 2 μM to about 50 μM within the composition. In some cases, Y-27632 is present at about 4 μM to about 25 μM within the composition. In some cases, Y-27632 is present at about 10 μM within the composition. In some cases, the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409. In some cases, the CBP/p300 bromodomain inhibitor comprises SGC-CBP30. In some cases, SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM within the composition. In some cases, SGC-CBP30 is present at about 0.4 μM to about 10 μM within the composition. In some cases, SGC-CBP30 is present at about 0.8 μM to about 5 μM within the composition. In some cases, SGC-CBP30 is present at about 2 μM within the composition. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469. In some cases, VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM within the composition. In some cases, VTP50469 is present at about 0.1 μM to about 2.5 μM within the composition. In some cases, VTP50469 is present at about 0.2 μM to about 1.25 μM with-in the composition. In some cases, VTP50469 is present at about 0.5 μM within the composition. In some cases, the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor. In some cases, AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM within the composition. In some cases, AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM within the composition. In some cases, AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM within the composition. In some cases, AKT Kinase Inhibitor is present at about 1 μM within the composition. In some cases, the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014. In some cases, the glycogen kinase inhibitor comprises CHIR99021. In some cases, CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM within the composition. In some cases, CHIR99021 is present at about 1 μM to about 25 μM within the composition. In some cases, CHIR99021 is present at about 2 μM to about 12.5 μM with-in the composition. In some cases, CHIR99021 is present at about 5 μM within the composition. In some cases, the TGFβ receptor inhibitor is an ALK5 inhibitor. In some cases, the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334, In some cases, the TGFβ receptor inhibitor comprises E-616452. In some cases, E-616452 is present at about 1 micromolar (μM) to about 100 μM within the composition. In some cases, E-616452 is present at about 2 μM to about 50 μM within the composition. In some cases, E-616452 is present at about 4 μM to about 25 μM within the composition. In some cases, E-616452 is present at about 10 μM within the composition. In some cases, the RAR agonist comprises TTNPB, Ch55, or AM580. In some cases, the RAR agonist comprises TTNPB. In some cases, TTNPB is present at about 0.2 μM to about 20 μM within the composition. In some cases, TTNPB is present at about 0.4 μM to about 10 μM within the composition. In some cases, TTNPB is present at about 0.8 μM to about 5 μM within the composition. In some cases, TTNPB is present at about 2 μM within the composition. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG. In some cases, SAG is present at about 0.05 micromolar (μM) to about 5 μM within the composition. In some cases, SAG is present at about 0.1 μM to about 2.5 μM within the composition. In some cases, SAG is present at about 0.2 μM to about 1.25 μM within the composition. In some cases, SAG is present at about 0.5 μM within the composition. In some cases, the Dot1L inhibitor comprises EPZ004777 or EPZ5676. In some cases, the Dot1L inhibitor comprises the EPZ5676. In some cases, EPZ5676 is present at about 0.2 μM to about 20 μM within the composition. In some cases, EPZ5676 is present at about 0.4 μM to about 10 μM within the composition. In some cases, EPZ5676 is present at about 0.8 μM to about 5 μM within the composition. In some cases, EPZ5676 is present at about 2 μM within the composition. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib. In some cases, Ruxolitinib is present at about 0.1 μM to about 10 μM within the composition. In some cases, Ruxolitinib is present at about 0.2 μM to about 5 μM within the composition. In some cases, Ruxolitinib is present at about 0.4 μM to about 2.5 μM with-in the composition. In some cases, Ruxolitinib is present at about 1 μM within the composition. In some cases, the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA. In some cases, the SAH hydrolase inhibitor comprises DZNep. In some cases, DZNep is present at about 0.02 μM to about 2 μM within the composition. In some cases, DZNep is present at about 0.04 μM to about 1 μM within the composition. In some cases, DZNep is present at about 0.08 μM to about 0.5 μM within the composition. In some cases, DZNep is present at about 0.2 μM with the composition. In some cases, the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12. In some cases, the c-Jun kinase inhibitor comprises JNKIN8. In some cases, JNKIN is present at about 0.05 micromolar (μM) to about 5 μM within the composition. In some cases, JNKIN is present at about 0.1 μM to about 2.5 μM within the composition. In some cases, JNKIN is present at about 0.2 μM to about 1.25 μM within the composition. In some cases, JNKIN is present at about 0.5 μM within the composition. In some cases, the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702. In some cases, the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) . In some cases, 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM within the composition. In some cases, 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM within the composition. In some cases, 5-ITU is present at about 0.2 micromolar μM to about 1 μM within the composition. In some cases, the 5-ITU is present at about 0.5 μM within the composition. In some cases, the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid. In some cases, the casein kinase 2 inhibitor comprises the CX-4945. In some cases, CX-4945 is present at about 0.1 μM about 10 μM within the composition. In some cases, CX-4945 is present at about 0.2 μM about 5 μM within the composition. In some cases, CX-4945 is present at about 0.5 μM about 1 μM within the composition. In some cases, the G9a inhibitor comprises Unc0224, Unc0638, Unc0642, or Bix01294. In some cases, the G9a inhibitor comprises Unc0224. In some cases, Unc0224 is present at about 0.1 μM to about 10 μM within the composition. In some cases, Unc0224 is present at about 0.2 μM to about 5 μM within the composition. In some cases, Unc0224 is present at about 0.5 μM to about 2 μM within the composition. In some cases, Unc0224 is present at about 1 μM within the composition. In some cases, the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027. In some cases, the DNMT inhibitor comprises 5-Azacytidine. In some cases, 5-Azacytidine is present at about 0.2 μM to about 20 μM within the composition. In some cases, 5-Azacytidine is present at about 0.5 μM to about 10 μM within the composition. In some cases, 5-Azacytidine is present at about 1 μM to about 5 μM within the composition. In some cases, 5-Azacytidine is present at about 2 μM within the composition. In some cases, the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190. In some cases, the p38 MAPK inhibitor comprises BIRB796. In some cases, BIRB796 is present at about 0.2 μM to about 20 μM within the composition. In some cases, BIRB796 is present at about 0.4 μM to about 10 μM within the composition. In some cases, BIRB796 is present at about 0.8 μM to about 5 μM within the composition. In some cases, BIRB796 is present at about 2 μM within the composition.

Provided herein, in some aspects, are compositions. In some aspects, a composition provided herein is a medium for culturing cells in vitro. In some aspects, a composition provided herein comprises: (1) one or more of a G9a inhibitor or a DNMT inhibitor; and (2) one or more of a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, an SAH hydrolase inhibitor, a c-Jun kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or a p38 MAPK inhibitor. In some cases, the composition comprises: the glycogen kinase inhibitor, the TGFβ receptor inhibitor, the retinoic acid receptor agonist, the ROCK inhibitor, the CBP/p300 bromodomain inhibitor, the Menin-MLL interaction inhibitor, the serine-threonine kinase Akt inhibitor, the agonist for the G protein-coupled receptor Smoothened, and the Dot1L inhibitor.

In some cases, the composition comprises the SAH hydrolase inhibitor, and the c-Jun kinase inhibitor. In some cases, the composition comprises the G9a inhibitor, the DNMT inhibitor, and the p38 MAPK inhibitor.

In some cases, the ROCK inhibitor comprises Y-27632 or thiazovivin. In some cases, the ROCK inhibitor comprises Y-27632. In some cases, Y-27632 is present at about 1 μM to about 100 μM in the composition. In some cases, Y-27632 is present at about 2 μM to about 50 μM in the composition. In some cases, Y-27632 is present at about 4 μM to about 25 μM in the composition. In some cases, Y-27632 is present at about 10 μM in the composition. In some cases, the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409. In some cases, the CBP/p300 bromodomain inhibitor comprises SGC-CBP30. In some cases, SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM in the composition wherein SGC-CBP30 is present at about 0.4 μM to about 10 μM in the composition. In some cases, SGC-CBP30 is present at about 0.8 μM to about 5 μM in the composition. In some cases, SGC-CBP30 is present at about 2 μM in the composition. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4. In some cases, the Menin-MLL interaction inhibitor comprises VTP50469. In some cases, VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM in the composition. In some cases, VTP50469 is present at about 0.1 μM to about 2.5 μM in the composition. In some cases, VTP50469 is present at about 0.2 μM to about 1.25 μM in the composition. In some cases, VTP50469 is present at about 0.5 μM in the composition. In some cases, the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor. In some cases, AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM in the composition. In some cases, AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM in the composition. In some cases, AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM in the composition. In some cases, AKT Kinase Inhibitor is present at about 1 μM in the composition. In some cases, the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014. In some cases, the glycogen kinase inhibitor comprises CHIR99021. In some cases, CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM in the composition. In some cases, CHIR99021 is present at about 1 μM to about 25 μM in the composition. In some cases, CHIR99021 is present at about 2 μM to about 12.5 μM in the composition. In some cases, CHIR99021 is present at about 5 μM in the composition. In some cases, the TGFβ receptor inhibitor is an ALK5 inhibitor. In some cases, the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334. In some cases, the TGFβ receptor inhibitor comprises E-616452. In some cases, E-616452 is present at about 1 micromolar (μM) to about 100 μM in the composition. In some cases, E-616452 is present at about 2 μM to about 50 μM in the composition. In some cases, E-616452 is present at about 4 μM to about 25 μM in the composition. In some cases, E-616452 is present at about 10 μM in the composition. In some cases, the RAR agonist comprises TTNPB, Ch55, or AM580. In some cases, the RAR agonist comprises TTNPB. In some cases, TTNPB is present at about 0.2 μM to about 20 μM in the composition. In some cases, TTNPB is present at about 0.4 μM to about 10 μM in the composition. In some cases, TTNPB is present at about 0.8 μM to about 5 μM in the composition. In some cases, TTNPB is present at about 2 μM in the composition. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH. In some cases, the agonist for the G protein-coupled receptor Smoothened comprises SAG. In some cases, SAG is present at about 0.05 micromolar (μM) to about 5 μM in the composition. In some cases, SAG is present at about 0.1 μM to about 2.5 μM in the composition. In some cases, SAG is present at about 0.2 μM to about 1.25 μM in the composition. In some cases, SAG is present at about 0.5 μM in the composition. In some cases, the Dot1L inhibitor comprises EPZ004777 or EPZ5676. In some cases, the Dot1L inhibitor comprises the EPZ5676. In some cases, EPZ5676 is present at about 0.2 μM to about 20 μM in the composition. In some cases, EPZ5676 is present at about 0.4 μM to about 10 μM in the composition. In some cases, EPZ5676 is present at about 0.8 μM to about 5 μM in the composition. In some cases, EPZ5676 is present at about 2 μM in the composition. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib. In some cases, the Jak1/Jak2 inhibitor comprises Ruxolitinib. In some cases, Ruxolitinib is present at about 0.1 μM to about 10 μM in the composition. In some cases, Ruxolitinib is present at about 0.2 μM to about 5 μM in the composition. In some cases, Ruxolitinib is present at about 0.4 μM to about 2.5 μM in the composition. In some cases, Ruxolitinib is present at about 1 μM in the composition. In some cases, the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA. In some cases, the SAH hydrolase inhibitor comprises DZNep. In some cases, DZNep is present at about 0.02 μM to about 2 μM in the composition. In some cases, DZNep is present at about 0.04 μM to about 1 μM in the composition. In some cases, DZNep is present at about 0.08 μM to about 0.5 μM in the composition. In some cases, DZNep is present at about 0.2 μM in the composition. In some cases, the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12. In some cases, the c-Jun kinase inhibitor comprises JNKIN8. In some cases, JNKIN is present at about 0.05 micromolar (μM) to about 5 μM in the composition. In some cases, JNKIN is present at about 0.1 μM to about 2.5 μM in the composition. In some cases, JNKIN is present at about 0.2 μM to about 1.25 μM in the composition. In some cases, JNKIN is present at about 0.5 μM in the composition. In some cases, the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702. In some cases, the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) . In some cases, 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM in the composition. In some cases, 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM in the composition. In some cases, 5-ITU is present at about 0.2 micromolar μM to about 1 μM in the composition. In some cases, the 5-ITU is present at about 0.5 μM in the composition. In some cases, the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid. In some cases, the casein kinase 2 inhibitor comprises the CX-4945. In some cases, CX-4945 is present at about 0.1 μM about 10 μM in the composition. In some cases, CX-4945 is present at about 0.2 μM about 5 μM in the composition. In some cases, CX-4945 is present at about 0.5 μM about 1 μM in the composition. In some cases, the G9a inhibitor comprises Unc0224, Unc0638, Unc0642 , or Bix01294. In some cases, the G9a inhibitor comprises Unc0224. In some cases, Unc0224 is present at about 0.1 μM to about 10 μM in the composition. In some cases, Unc0224 is present at about 0.2 μM to about 5 μM in the composition. In some cases, Unc0224 is present at about 0.5 μM to about 2 μM in the composition. In some cases, Unc0224 is present at about 1 μM in the composition. In some cases, the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027. In some cases, the DNMT inhibitor comprises 5-Azacytidine. In some cases, 5-Azacytidine is present at about 0.2 μM to about 20 μM in the composition. In some cases, 5-Azacytidine is present at about 0.5 μM to about 10 μM in the composition. In some cases, 5-Azacytidine is present at about 1 μM to about 5 μM in the composition. In some cases, 5-Azacytidine is present at about 2 μM in the composition. In some cases, the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190. In some cases, the p38 MAPK inhibitor comprises BIRB796. In some cases, BIRB796 is present at about 0.2 μM to about 20 μM in the composition. In some cases, BIRB796 is present at about 0.4 μM to about 10 μM in the composition. In some cases, BIRB796 is present at about 0.8 μM to about 5 μM in the composition. In some cases, BIRB796 is present at about 2 μM in the composition.

Provided herein, in some aspects, is a composition comprising: (a) somatic cells, and (b) a c-Jun kinase inhibitor or a CBP/p300 bromodomain inhibitor. In some cases, the composition further comprises a serine-threonine kinase Akt inhibitor.

Provided herein, in some aspects, is a composition comprising: (a) somatic cells, and (b) one or more of a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor.

In some cases, the composition further comprises a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a Menin-MLL interaction inhibitor, or both. In some cases, the composition further comprises one or more of: (1) a glycogen kinase inhibitor; (2) a TGFβ receptor inhibitor; (3) a retinoic acid receptor agonist; (4) an agonist for the G protein-coupled receptor Smoothened; (5) a Dot1L inhibitor; (6) a Jak1/Jak2 inhibitor; or (7) an SAH hydrolase inhibitor. 397. The composition of any one of claims 392-396, wherein the composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, a p38 MAPK inhibitor. In some cases, the composition further comprises a G9a inhibitor or a DNMT inhibitor. In some cases, the composition comprises the adenosine kinase inhibitor, the casein kinase 2 inhibitor, or both. In some cases, the somatic cells comprise primary hu-man adult adipose derived mesenchymal stromal cells (hADSCs) . In some cases, the somatic cells comprise fibroblasts.

In some cases, the somatic cells comprise a genetic modification. In some cases, the genetic modification comprises an exogenous nucleic acid sequence. In some cases, the exogenous nucleic acid sequence encodes a polypeptide. In some cases, the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule. In some cases, the genetic modification comprises alteration of a genomic sequence. In some cases, the genetic modification reduces immunogenicity of the somatic cells.

Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “FIG. ” or “FIGs. ” herein) , of which:

FIG. 1 depicts a schematic of an exemplary method for chemically inducing human somatic cells into human chemically induced pluripotent stem cells (hCiPSCs) .

FIG. 2 shows a picture of a OCT4-positive human chemically induced pluripotent stem cells (hCiPSCs) colony induced by a two-stage protocol according to some embodiments of the present disclosure.

FIG. 3 depicts gene expression profile of exemplary intermediate plastic state cells in stage 2.

FIG. 4 depicts exemplary compositions comprising chemical reprogramming factors and optional cells in stage 1.

FIG. 5 depicts exemplary CiPSCs that express OCT4, SOX2, and NANOG shown in a heatmap.

FIG. 6 depicts exemplary compositions comprising chemical reprogramming factors and optional cells in stage 2.

DETAILED DESCRIPTION

Definitions

The term “converting” or “reprogramming, ” and their grammatical equivalents as used herein when referring to cells refers to a process that alters or reverses the differentiation state of a cell (e.g., a somatic cell) . As used herein, a conversion process, when referring to conversion of a cell of a first cell type into a cell of a second cell type, refer to a process where the cell of the first cell type is converted into a cell of the second cell type, or a process where one or more progenies of the cell of the first cell type is converted into a cell of the second cell type.

The term “differentiation” and its grammatical equivalents as used herein can refer to the process by which a less specialized cell (e.g., a more naive cell with a higher cell potency) becomes a more specialized cell type (e.g., a less naive cell with a lower cell potency) ; and that the term “de-differentiation” can refer to the process by which a more specialized cell becomes a less specialized cell type (e.g., a more naive cell with a higher cell potency) .

As used herein, the term “cell potency” can refer to the ability of a cell to differentiate into cells of different lineages. For example, without wishing to be bound by a certain theory, a pluripotent cell (e.g., a stem cell) has the potential to differentiate into cells of, or derived from, any of the three germ layers, that is, endoderm (e.g., interior stomach lining, gastrointestinal tract, the lungs) , mesoderm (e.g., muscle, bone, blood, urogenital) , or ectoderm (e.g., epidermal tissues and nervous system) , and accordingly has high cell potency; a multipotent cell (e.g., a stem cell of a certain type, such as hematopoietic stem cells, cardiac stem cells, or neural stem cells, etc. ) has the ability to give rise to cells from a multiple, but limited, number of lineages (such as blood cell lineage, cardiac cell lineage, neural cell lineage) comparatively has a lower cell potency than pluripotent cells. Cells that are committed to a particular lineage or are terminally differentiated can have yet a lower cell potency.

The term “isolated population of cells” or “isolated cell population” as used herein refers to a group of non-naturally occurring cells.

The term “population of cells, ” “cell population, ” or grammatically equivalent thereof, as used herein refers to a group of cells, their progenies, or progenies thereof, and/or the cells derived from thereof. For example, a population of first cells may comprise the first cells or the progenies of the first cells.

The term “progenies, ” when used herein with reference toa cell, can refer to any of the daughter cells derived from mitotic division of the cell or mitotic division of any of the progenies of the cell.

Whenever the term “at least, ” “greater than, ” or “greater than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “at least” or “greater than” applies to each one of the numerical values in that series of numerical values.

Whenever the term “at most, ” “no more than, ” “less than, ” or “less than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “no more than” or “less than” applies to each one of the numerical values in that series of numerical values.

As used herein, the singular forms “a” , “an” , and “the” include plural references unless the context clearly dictates otherwise.

The term “and/or” as used in a phrase such as “A and/or B” herein is intended to include both A and B; A or B; A (alone) ; and B (alone) . Likewise, the term "and/or" as used in a phrase such as “A, B, and/or C” is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone) ; B (alone) ; and C (alone) .

The term “about” or “approximately” as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1%of the specified amount. For example, “about” can mean plus or minus 10%, per the practice in the art. Alternatively, “about” can mean a range of plus or minus 20%, plus or minus 10%, plus or minus 5%, or plus or minus 1%of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, up to 5-fold, or up to 2-fold, of a value. Where particular values can be described in the application and claims, unless otherwise stated the term “about” meaning up to an acceptable error range for the particular value should be assumed. Also, where ranges, subranges, or both, of values can be provided, the ranges or subranges can include the endpoints of the ranges or subranges. The terms “substantially” , “substantially no” , “substantially free” , and “approximately” can be used when describing a magnitude, a position or both to indicate that the value described can be up to a reasonable expected range of values. For example, a numeric value can have a value that can be +/-0.1%of the stated value (or range of values) , +/- 1%of the stated value (or range of values) , +/-2%of the stated value (or range of values) , +/-5%of the stated value (or range of values) , +/-10%of the stated value (or range of values) , etc. Any numerical range recited herein can be intended to include all sub-ranges subsumed therein.

METHODS

Provided herein, in some aspects, are methods and compositions for cell conversions. In some instances, a method may comprise contacting a first population cells with a composition comprising one or more reprogramming factors. In some case, the contacting may comprise incubating a first population of cells with a composition comprising one or more reprogramming factors for a period of time. Subsequent to the contacting, at least a subset of the first population of cells may be converted into different cells. The population of cells subsequent to the conversion may comprise a second population of cells. During or subsequent to the conversion, the first population and second population of cells may have different biological properties. The different biological properties may comprise different expressions of genes, different expression of proteins, different cell proliferation properties (e.g., cell divisions or cell growth/increase of cell masses) , different sizes of cells, different numbers of cells, different modifications of the genomes of the cells (e.g., epigenetic modifications) , different cell cycle stages, different senescence stages, or different differentiation stages or types, or any combination thereof. The first population and second population of cells may differ by having at least two cells that have at least a different cellular activity. The first population and second population of cells may differ by having different cell types. The different cell types may have different cellular activities described herein.

Provided herein, in some aspects, are methods and compositions for converting cells in at most 2 stages. The methods and compositions for converting cells may comprise 2 stages. The 2 stages may comprise stage 1 (or stage I) and stage 2 (or stage II) . The methods and compositions described herein may comprise stage 1 that involves conversion of a somatic cell into a cell with a higher cell potency (e.g., less specialized cell) , such as an intermediate plastic state cell. The methods and compositions described herein may comprise stage 2 that involves conversion of an intermediate plastic state cell into a cell with a pluripotent stem cell. A stage of the plurality of stages may comprise contacting a first population of cells with a first composition and converting at least a subset of the first population of cells into different cells. The population of cells subsequent to the conversion may comprise a second population of cells.

In some cases, a method may comprise (1) contacting a first population of cells with a first composition; (2) converting at least a subset of the first population of cells into different cells and generating a second population of cells comprising the converted cells; (3) contacting the second population of cells with a second composition; (4) converting at least a subset of the second population of cells into different cells and generating a third population of cells comprising the converted cells. The first and second compositions may each comprise one or more reprogramming factors. Any of the first and second composition may comprise one or more reprogramming factors. The first and second compositions may be different. The first, second, and third populations of cells may be different. In some cases, prior to, during, or subsequent to each contacting, at least a cell of the population of cells being contacted with a composition may be cultured. In some cases, prior to, during, or subsequent to each converting, the population of cells being contacted with a composition may be cultured. When the cells are cultured, the cells may undergo cell proliferation.

It will be appreciated that the ordinal numbering of a populations of cells or composition relative to other populations of cells or compositions may not limit the populations of cells or composition to specific populations of cells or composition. In some cases, the ordinal numbering of a populations of cells or composition should be understood to distinguish a population of cells or compositions from another population (s) of cells or composition (s) .

In some cases, a method may comprise (1) contacting a first population of cells comprising somatic cells with a first composition; (2) converting at least a subset of the somatic cells or the progenies thereof into different cells and generating a second population of cells comprising the converted cells comprising intermediate plastic state cells; (3) contacting the second population of cells comprising intermediate plastic state cells with a second composition; (4) converting at least a subset of the second population of cells into different cells and generating a third population of cells comprising the converted cells comprising pluripotent stem cells.

In some cases, a method may convert a population of cells comprising somatic cells into a different population of cells comprising pluripotent stem cells within a pluripotent stem cell conversion time period. The pluripotent stem cell conversion time period may comprise the time period from contacting the population of cells comprising somatic cells to the time when at least a pluripotent stem cell is generated. The pluripotent stem cell conversion time period may be less than about 50 days, 49 days, 48 days, 47 days, 46 days, 45 days, 44 days, 43 days, 42 days, 41 days, 40 days, 39 days, 38 days, 37 days, 36 days, 35 days, 34 days, 33 days, 32 days, 31 days, 30 days, 29 days, 28 days, 27 days, 26 days, 25 days, 24 days, 23 days, 22 days, 21 days, 20 days, 19 days, 18 days, 17 days, 16 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days or less. The pluripotent stem cell conversion time period may be less than about 40 days. The pluripotent stem cell conversion time period may be less than about 39 days. The pluripotent stem cell conversion time period may be less than about 38 days. The pluripotent stem cell conversion time period may be less than about 37 days. The pluripotent stem cell conversion time period may be less than about 36 days. The pluripotent stem cell conversion time period may be less than about 35 days. The pluripotent stem cell conversion time period may be less than about 34 days. The pluripotent stem cell conversion time period may be less than about 33 days. The pluripotent stem cell conversion time period may be less than about 32 days. The pluripotent stem cell conversion time period may be less than about 31 days. The pluripotent stem cell conversion time period may be less than about 30 days. The pluripotent stem cell conversion time period may be less than about 29 days. The pluripotent stem cell conversion time period may be less than about 28 days. The pluripotent stem cell conversion time period may be less than about 27 days. The pluripotent stem cell conversion time period may be less than about 26 days. The pluripotent stem cell conversion time period may be less than about 25 days. The pluripotent stem cell conversion time period may be less than about 24 days. The pluripotent stem cell conversion time period may be less than about 23 days. The pluripotent stem cell conversion time period may be less than about 22 days. The pluripotent stem cell conversion time period may be less than about 21 days. The pluripotent stem cell conversion time period may be less than about 20 days. The pluripotent stem cell conversion time period may be less than about 19 days. The pluripotent stem cell conversion time period may be less than about 18 days. The pluripotent stem cell conversion time period may be less than about 17 days. The pluripotent stem cell conversion time period may be less than about 16 days. The pluripotent stem cell conversion time period may be less than about 15 days.

The pluripotent stem cell conversion time period may be about 32 days. The pluripotent stem cell conversion time period may be about 31 days. The pluripotent stem cell conversion time period may be about 30 days. The pluripotent stem cell conversion time period may be about 29 days. The pluripotent stem cell conversion time period may be about 28 days. The pluripotent stem cell conversion time period may be about 27 days. The pluripotent stem cell conversion time period may be about 26 days. The pluripotent stem cell conversion time period may be about 25 days. The pluripotent stem cell conversion time period may be about 24 days. The pluripotent stem cell conversion time period may be about 23 days. The pluripotent stem cell conversion time period may be about 22 days. The pluripotent stem cell conversion time period may be about 21 days. The pluripotent stem cell conversion time period may be about 20 days. The pluripotent stem cell conversion time period may be about 19 days. The pluripotent stem cell conversion time period may be about 18 days. The pluripotent stem cell conversion time period may be about 17 days. The pluripotent stem cell conversion time period may be about 16 days. The pluripotent stem cell conversion time period may be about 15 days.

In some cases, a method may convert a population of cells comprising somatic cells into a different population of cells comprising pluripotent stem cells with a pluripotent stem cell conversion efficiency. The pluripotent stem cell conversion efficiency may be measured as a ratio (e.g., percentage) of the number of pluripotent stem cells generated relative to the number of somatic cells, to which the method disclosed herein is applied for generating the number of pluripotent stem cells. For example, if the method generates one pluripotent stem cell from 1000 somatic cells, the pluripotent stem cell conversion efficiency of the method is 0.1%. In some cases, the pluripotent stem cell conversion efficiency of the method may be at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.5%or more. In some cases, the pluripotent stem cell conversion efficiency of the method may be about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, or 0.5%. The pluripotent stem cell conversion efficiency of the method may be about 0.0005%. The pluripotent stem cell conversion efficiency of the method may be about 0.0006%. The pluripotent stem cell conversion efficiency of the method may be about 0.0007%. The pluripotent stem cell conversion efficiency of the method may be about 0.0008%. The pluripotent stem cell conversion efficiency of the method may be about 0.0009%. The pluripotent stem cell conversion efficiency of the method may be about 0.001%. The pluripotent stem cell conversion efficiency of the method may be about 0.002%. The pluripotent stem cell conversion efficiency of the method may be about 0.003%. The pluripotent stem cell conversion efficiency of the method may be about 0.004%. The pluripotent stem cell conversion efficiency of the method may be about 0.005%. The pluripotent stem cell conversion efficiency of the method may be about 0.006%. The pluripotent stem cell conversion efficiency of the method may be about 0.007%. The pluripotent stem cell conversion efficiency of the method may be about 0.008%. The pluripotent stem cell conversion efficiency of the method may be about 0.009%. The pluripotent stem cell conversion efficiency of the method may be about 0.01%. The pluripotent stem cell conversion efficiency of the method may be about 0.02%. The pluripotent stem cell conversion efficiency of the method may be about 0.03%. The pluripotent stem cell conversion efficiency of the method may be about 0.04%. The pluripotent stem cell conversion efficiency of the method may be about 0.05%. The pluripotent stem cell conversion efficiency of the method may be about 0.06%. The pluripotent stem cell conversion efficiency of the method may be about 0.07%. The pluripotent stem cell conversion efficiency of the method may be about 0.08%. The pluripotent stem cell conversion efficiency of the method may be about 0.09%. The pluripotent stem cell conversion efficiency of the method may be about 0.1%.

In some cases, provided herein is a method for producing pluripotent stem cells. In some cases, the method comprises contacting somatic cells with a first composition, thereby obtaining a first cell population. In some cases, the first composition contacted to the somatic cells comprises one or more of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor; a CBP/p300 bromodomain inhibitor; a Menin-MLL interaction inhibitor; a serine-threonine kinase Akt inhibitor; a glycogen kinase inhibitor; a TGFβ receptor inhibitor; a retinoic acid receptor agonist; an agonist for the G protein-coupled receptor Smoothened; a Dot1L inhibitor; a Jak1/Jak2 inhibitor; an SAH hydrolase inhibitor; or a c-Jun kinase inhibitor. In some cases, the method further comprises contacting the first cell population with a second composition, thereby obtaining a second cell population comprising pluripotent stem cells. In some cases, the second composition contacted to the first cell population comprises one or more of a MEK inhibitor; a B-Raf inhibitor; or a histone deacetylase inhibitor.

In some cases, the first composition contacted to the somatic cells comprises one or more of the c-Jun kinase inhibitor, the CBP/p300 bromodomain inhibitor, or the serine-threonine kinase Akt inhibitor. In some cases, the first composition contacted to the somatic cells comprises the c-Jun kinase inhibitor, the CBP/p300 bromodomain inhibitor, and the serine-threonine kinase Akt inhibitor. In some cases, the first composition contacted to the somatic cells comprises one or more of the ROCK inhibitor; the CBP/p300 bromodomain inhibitor; the Menin-MLL interaction inhibitor; or the serine-threonine kinase Akt inhibitor. In some cases, the first composition contacted to the somatic cells comprises one or more of the ROCK inhibitor, the c-Jun kinase inhibitor, or the CBP/p300 bromodomain inhibitor. In some cases, the first composition comprises the Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor; the CBP/p300 bromodomain inhibitor; the Menin-MLL interaction inhibitor; the serine-threonine kinase Akt inhibitor; the glycogen kinase inhibitor; the TGFβ receptor inhibitor; the retinoic acid receptor agonist; an agonist for the G protein-coupled receptor Smoothened; the Dot1L inhibitor; the Jak1/Jak2 inhibitor; the SAH hydrolase inhibitor; and the c-Jun kinase inhibitor. In some cases, the first composition contacted to the somatic cells further comprises one or more an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNA methyltransferase (DNMT) inhibitor, or a p38 MAPK inhibitor. In some cases, the first composition contacted to the somatic cells further comprises a G9a inhibitor or a DNMT inhibitor. In some cases, the first composition contacted to the somatic cells further comprises an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or both.

In some cases, the second composition contacted to the first cell population further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor. In some cases, the second composition contacted to the first cell population further comprises one or more of an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor. In some cases, the second composition contacted to the first cell population further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, or a ROCK inhibitor.

In some cases, the somatic cells comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) . In some cases, the somatic cells comprise fibroblasts. The somatic cells may comprise a genetic modification. In some cases, the pluripotent stem cells comprise the genetic modification. The genetic modification may comprise an exogenous nucleic acid sequence. In some cases, the exogenous nucleic acid sequence encodes a polypeptide. In some cases, the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule. The genetic modification may comprise alteration of a genomic sequence. The genetic modification may reduce immunogenicity of the pluripotent stem cells. Reduced immunogenicity of the pluripotent stem cells may help improve the therapeutic effects and prevent or reduce side effects (such as immune response from the host) of a cell therapy involving administration of the pluripotent stem cells or cell derived therefrom.

A cell in any populations of cells described herein may comprise a mammalian cell. A cell may comprise a mouse cell, a hamster cell, a rat cell, or a rodent cell. A cell may comprise a mouse cell. A cell may comprise a hamster cell. A cell may comprise a rat cell. A cell may comprise a rodent cell. In some cases, a cell may comprise a primate cell. A cell may comprise a strepsirrhine cell or a haplorrhine cell. A cell may comprise a monkey cell, an ape cell, or a human cell. In some cases, a cell may comprise a human cell. A cell may comprise a monkey cell. A cell may comprise an ape cell.

STAGE 1

In some aspects, provided herein are methods and compositions for conversion of a somatic cell into a cell with a higher cell potency (e.g., less specialized cell) , such as an intermediate plastic state cell-the conversion process referred herein also as "stage 1. " A stage 1 method may be part of a conversion process that reprograms somatic cells into pluripotent stem cells. A stage 1 method may be the first stage of a conversion process that reprograms somatic cells into pluripotent stem cells.

A stage 1 method may comprise contacting a first cell population with a first composition. A stage 1 method may comprise, subsequent to or during the contacting, converting a subset of the first cell population into different cells. The cell population comprising the different cells may comprise a second cell population. A stage 1 method may comprise incubating the first cell population with the first composition for a period of time. The subset of the first cell population may be converted into the different cells prior to, during, or subsequent to the incubating. In some cases, a stage 1 method may comprise removing the first composition from the second population of cells. In other cases, a stage 1 method may comprise removing the first composition from the first population of cells.

The first population of stage 1 cells may comprise somatic cells. A somatic cell may comprise a skin cell, a nerve cell, a muscle cell, or a blood cell. A somatic cell may not comprise a germ cell. A somatic cell may not comprise an undifferentiated cell. A somatic cell may not comprise a gamete (sperms or eggs) . A somatic cell may not comprise a gametocyte. In some cases, a somatic cell may also comprise a muscle cell, a fat cell, a connective tissue cell, a vasculature cell, a neuron, a bone cell, or a skin cell. The first population of stage 1 cells may comprise fibroblasts, primary human adult adipose derived mesenchymal stromal cells (hADSCs) , smooth muscle cells, cardiac muscle cells, skeletal muscle cells, neurons, red blood cells, white blood cells, platelets, osteoblasts, osteoclasts, squamous cells, basal cells, or melanocytes, or any combination thereof. The first population of stage 1 cells may comprise fibroblasts or hADSCs. The first population of stage 1 cells may comprise fibroblasts. The first population of stage 1 cells may comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) . The first population of stage 1 cells may comprise smooth muscle cells. The first population of stage 1 cells may comprise cardiac muscle cells. The first population of stage 1 cells may comprise skeletal muscle cells. The first population of stage 1 cells may comprise neurons. The first population of stage 1 cells may comprise red blood cells. The first population of stage 1 cells may comprise white blood cells. The first population of stage 1 cells may comprise platelets. The first population of stage 1 cells may comprise osteoblasts. The first population of stage 1 cells may comprise osteoclasts. The first population of stage 1 cells may comprise squamous cells. The first population of stage 1 cells may comprise basal cells. The first population of stage 1 cells may comprise melanocytes. The first population of stage 1 cells may comprise cells from the intestinal epithelium. The first population of stage 1 cells may comprise neonatal (for example foreskin) or adult fibroblasts. The first population of stage 1 cells may comprise epithelial cells, endothelial cells, cells of mesenchymal origin, parenchymal cells (for example, hepatocytes) , neurological cells, or connective tissue cells, or any combination thereof. The first population of stage 1 cells may not comprise cells that are not somatic cells. In some cases, the first population of stage 1 cells may comprise germ cells.

The first population of stage 1 cells may be isolated by disaggregating an appropriate organ or tissue. For example, the tissue or organ can be disaggregated mechanically and/or treated with digestive enzymes and/or chelating agents that weaken the connections between neighboring cells, so that the tissue can be dispersed to form a suspension of individual cells without appreciable cell breakage. Enzymatic dissociation can be accomplished by mincing the tissue and treating the minced tissue with one or more enzymes such as trypsin, chymotrypsin, collagenase, elastase, and/or hyaluronidase, DNase, pronase, dispase etc. Mechanical disruption can also be accomplished by a number of methods including, but not limited to, the use of grinders, blenders, sieves, homogenizers, pressure cells, or insonators.

The second population of stage 1 cells may comprise intermediate plastic state cells.The first population of stage 1 cells may not comprise intermediate plastic state cells. An intermediate plastic state cell may not be a naturally occurring cell. An intermediate plastic state cell may express a combination of genes that are not expressed by a naturally occurring cell. An intermediate plastic state cell may express at least one gene at level at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 5-fold, 100-fold, or higher, relative to a naturally occurring cell. An intermediate plastic state cell may express at least one gene at level at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%lower, relative to a naturally occurring cell. The second population of stage 1 cells may comprise somatic cells or intermediate plastic state cells. The second population of stage 1 cells may comprise somatic cells and intermediate plastic state cells. The second population of stage 1 cells may not comprise somatic cells. In some cases, the second population of stage 1 cells may comprise fewer somatic cells than the first population of stage 1 cells. For example, the second population of stage 1 cells may have 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%or 99% fewer somatic cells than the first population of stage 1 cells. In some cases, the second population of stage 1 cells may comprise more intermediate plastic state cells than the first population of stage 1 cells. For example, the second population of stage 1 cells may have 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 5-fold, 100-fold, or more intermediate plastic state cells than the first population of stage 1 cells.

Intermediate plastic state cells may show decreased expression of genes expressed by somatic cells. Intermediate plastic state cells may show increased expressions of genes involved in embryonic development, increased cell proliferation, and decreased methylation epigenetic state. Promoter regions of genes related to embryonic development, cell cycle and stem cell proliferation can be demethylated in intermediate plastic state cells. Intermediate plastic state cells may undergo dedifferentiation, relative to somatic cells. In some cases, genes related to limb and appendage development may be upregulated and have open chromatin structures in intermediate plastic state cells. Intermediate plastic state cells can be reprogrammed to acquire characteristics of developing human limb bud cells, similar to the situation of axolotl limb regeneration in which genes governing embryonic limb development are reactivated during dedifferentiation. However, dedifferentiation was not found in frogs and mice, according to Guan 2002, of which the limb tissue showed no notable activation of an embryonic gene expression program following injury.

An intermediate plastic state cell may express LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof. An intermediate plastic state cell may express LIN28A. An intermediate plastic state cell may express SALL4. An intermediate plastic state cell may express MSX2. An intermediate plastic state cell may express NMYC. An intermediate plastic state cell may express WNT4. An intermediate plastic state cell may express FGF19. An intermediate plastic state cell may express TOP2A. An intermediate plastic state cell may express MSX1. An intermediate plastic state cell may express HOXB9. An intermediate plastic state cell may express WT1. An intermediate plastic state cell may express GATA2. An intermediate plastic state cell may express HMGA2. An intermediate plastic state cell may express LEF1. An intermediate plastic state cell may express FGF9. An intermediate plastic state cell may express HOXA9. An intermediate plastic state cell may express HOZXA1. An intermediate plastic state cell may express PTCH1. An intermediate plastic state cell may express HOXA5. An intermediate plastic state cell may express CCND2. An intermediate plastic state cell may express SDC1. An intermediate plastic state cell may express TBX3. An intermediate plastic state cell may express BMP4. An intermediate plastic state cell may express IGF2. An intermediate plastic state cell may express one or more of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

An intermediate plastic state cell may express LIN28A or SALL4. An intermediate plastic state cell may express LIN28A and SALL4. An intermediate plastic state cell may express LIN28A and SALL4; and MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof. An intermediate plastic state cell may express LIN28A and SALL4; and MSX2. An intermediate plastic state cell may express LIN28A and SALL4; and NMYC. An intermediate plastic state cell may express LIN28A and SALL4; and WNT4. An intermediate plastic state cell may express LIN28A and SALL4; and FGF19. An intermediate plastic state cell may express LIN28A and SALL4; and TOP2A. An intermediate plastic state cell may express LIN28A and SALL4; and MSX1. An intermediate plastic state cell may express LIN28A and SALL4; and HOXB9. An intermediate plastic state cell may express LIN28A and SALL4; and WT1. An intermediate plastic state cell may express LIN28A and SALL4; and GATA2. An intermediate plastic state cell may express LIN28A and SALL4; and HMGA2. An intermediate plastic state cell may express LIN28A and SALL4; and LEF1. An intermediate plastic state cell may express LIN28A and SALL4; and FGF9. An intermediate plastic state cell may express LIN28A and SALL4; and HOXA9. An intermediate plastic state cell may express LIN28A and SALL4; and HOZXA1. An intermediate plastic state cell may express LIN28A and SALL4; and PTCH1. An intermediate plastic state cell may express LIN28A and SALL4; and HOXA5. An intermediate plastic state cell may express LIN28A and SALL4; and CCND2. An intermediate plastic state cell may express LIN28A and SALL4; and SDC1. An intermediate plastic state cell may express LIN28A and SALL4; and TBX3. An intermediate plastic state cell may express LIN28A and SALL4; and BMP4. An intermediate plastic state cell may express LIN28A and SALL4; and IGF2. An intermediate plastic state cell may express LIN28A and SALL4; and one or more of MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

An intermediate plastic state cell may express LIN28A and SALL4; a second gene; and a third gene. The second gene expressed by the intermediate plastic state cell may comprise MSX2, NMYC, WNT4, FGF19, or TOP2A, or any combination thereof. A somatic cell may not express SALL4. A somatic cell may not express both SALL4 and LIN28A. A somatic cell may not express SALL4; a second gene; or a third gene. A somatic cell may not express the second gene. A somatic cell may not express the third gene. A somatic cell may not express LIN28A or SALL4; may not express one or more second genes; and may not express one or more third genes. The second genes may comprise MSX2, NMYC, WNT4, FGF19, or TOP2A, or any combination thereof. The second genes may comprise MSX2. The second gene thereof may comprise NMYC. The second genes may comprise WNT4. The second gene thereof may comprise FGF19. The second genes may comprise TOP2A. The third genes may comprise MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1, or any combination thereof. The third genes may comprise MSX1. The third genes may comprise HOXB9. The third gene thereof may comprise WT1. The third genes may comprise GATA2. The third genes may comprise HMGA2. The third genes may comprise LEF1. An intermediate plastic state cell may express LIN28A and SALL4; a second gene; a third gene; and a fourth gene. A somatic cell may not express the fourth gene. The fourth gene may comprise FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof. The fourth gene may comprise FGF9. The fourth gene may comprise HOXA9. The fourth gene may comprise HOZXA1. The fourth gene may comprise PTCH1. The fourth gene may comprise HOXA5. The fourth gene may comprise CCND2. The fourth gene may comprise SDC1. The fourth gene may comprise TBX3. The fourth gene may comprise BMP4. The fourth gene may comprise IGF2.

A first population of stage 1 cells may not comprise the intermediate plastic state cell described herein. A somatic cell of the first population of stage 1 cells may not express LIN28A, NMYC, WNT2B, PAX8, SMAD3, GLI3, KRT18, KRT19, WT1, or TBX2, or any combination thereof. The somatic cell may not express LIN28A. The somatic cell may not express NMYC. The somatic cell may not express WNT2B. The somatic cell may not express PAX8. The somatic cell may not express SMAD3. The somatic cell may not express GLI3. The somatic cell may not express KRT18. The somatic cell may not express KRT19. The somatic cell may not express WT1. The somatic cell may not express TBX2. The somatic cell may not express LIN28A and may not one or more of NMYC, WNT2B, PAX8, SMAD3, GLI3, KRT18, KRT19, WT1, or TBX2. The somatic cell may not express LIN28A or NMYC. The somatic cell may not express LIN28A or WNT2B. The somatic cell may not express LIN28A or PAX8. The somatic cell may not express LIN28A or SMAD3. The somatic cell may not express LIN28A or GLI3. The somatic cell may not express LIN28A or KRT18. The somatic cell may not express LIN28A or KRT19. The somatic cell may not express LIN28A or WT1. The somatic cell may not express LIN28A or TBX2. The somatic cell may express MMP1, ZEB1, VIM, COL1A1, COL5A1, COL6A2, PRRX1, SNAI2, TWIST1, or TWIST2 or any combination thereof. The somatic cell may express MMP1. The somatic cell may express ZEB1. The somatic cell may express VIM. The somatic cell may express COL1A1. The somatic cell may express COL5A1. The somatic cell may express COL6A2. The somatic cell may express PRRX1. The somatic cell may express SNAI2. The somatic cell may express TWIST1. The somatic cell may express TWIST2.

FIG. 3 depicts exemplary intermediate plastic state cells in stage 2. In FIG. 3, "A" represents expression of LIN28A and SALL4; "B" represents expression of the second gene including MSX2, NMYC, WNT4, FGF19, or TOP2A, or any combination thereof; "C" represents expression the third gene including of MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1, or any combination thereof; "D" represents expression of the fourth gene including FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof. For example, AB1C1D1 represents intermediate plastic state cells that express LIN28A, SALL4, MSX2, and MSX1; AB1C1 represents intermediate plastic state cells that express LIN28A, SALL4, MSX2, MSX1, and FGF9. The cells can express more than one of B, C, or D. For example, AB1B2C1C2D1D2 represents intermediate plastic state cells that express LIN28A, SALL4, MSX2, NMYC, MSX1, HOXB9, FGF9 and HOXA9.

A cell of the second population of stage 1 cells may express higher levels of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof, relative to a cell of the first population of stage 1 cells. The higher level of expression of any one of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2 in a cell of the second population of stage 1 cells may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 5-fold, 100-fold, or more, relative to a cell of the first population of stage 1 cells. A cell of the first population of stage 1 cells may express lower levels of any one of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, relative to a cell of the second population of stage 1 cells. The lower level of expression of any one of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2 in a cell of the first population of stage 1 cells may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%, relative to a cell of the second population of stage 1 cells. The levels of expression can be measured by any methods described herein. For examples, gene expression can be measured by methods described in EXAMPLE 2. Gene expression can be measured by using any one of SEQ ID NO: 1-83 (including controls) .

In some aspects, provided herein is a composition that comprises reprogramming factors for stage 1 conversion, or comprises cells of stage 1 (the first population of cells or the second population of cells) , or comprises cells of stage 1 (the first population of cells or the second population of cells) and reprogramming factors for stage 1 conversion. In some cases, a composition comprises a culture medium comprising the reprogramming factors for stage 1 conversion.

In some cases, a composition may comprise an isolated population of the second population of stage 1 cells. In some cases, a composition may comprise an isolated population of the first population of stage 1 cells. An isolated population of stage 1 cells may comprise at least about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more cells. An isolated population of stage 1 cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 cells. An isolated population of stage 1 cells may comprise at least one intermediate plastic state cell. In some cases, an isolated population of stage 1 cells may comprise at least about 1, 1 x 10^1, 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more intermediate plastic state cells. An isolated population of cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 intermediate plastic state cells. In some cases, an isolated population of stage 1 cells may comprise at least about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more somatic cells. An isolated population of cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 somatic cells. In some cases, an isolated population of stage 1 cells may comprise at least about 1 x 10^1, 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more somatic cells, intermediate plastic state cells, or a combination thereof. An isolated population of stage 1 cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 intermediate plastic state cells, somatic cells, or a combination thereof.

A composition may comprise a chemical reprogramming factor. In some cases, the composition is a culture medium comprising one or more chemical reprogramming factors provided herein. A composition may comprise a plurality of chemical reprogramming factors. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chemical reprogramming factors. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chemical reprogramming factors. A composition may comprise 1 chemical reprogramming factors. A composition may comprise 2 chemical reprogramming factors. A composition may comprise 3 chemical reprogramming factors. A composition may comprise 4 chemical reprogramming factors. A composition may comprise 5 chemical reprogramming factors. A composition may comprise 6 chemical reprogramming factors. A composition may comprise 7 chemical reprogramming factors. A composition may comprise 8 chemical reprogramming factors. A composition may comprise 9 chemical reprogramming factors. A composition may comprise 10 chemical reprogramming factors. A composition may comprise 11 chemical reprogramming factors. A composition may comprise 12 chemical reprogramming factors. A composition may comprise 13 chemical reprogramming factors. A composition may comprise 14 chemical reprogramming factors. A composition may comprise 15 chemical reprogramming factors. A composition may comprise 16 chemical reprogramming factors. A composition may comprise 17 chemical reprogramming factors. A composition may comprise 18 chemical reprogramming factors. A composition may comprise 19 chemical reprogramming factors. A composition may comprise 20 chemical reprogramming factors. A chemical reprogramming factor in a composition may comprise any chemical reprogramming factors described here.

A composition may comprise a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor, or any combination thereof. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor.

A composition may comprise a Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor. A composition may comprise a CBP/p300 bromodomain inhibitor. A composition may comprise a Menin-MLL interaction inhibitor. A composition may comprise a serine-threonine kinase Akt inhibitor. A composition may comprise a glycogen kinase inhibitor. A composition may comprise a TGFβ receptor inhibitor. A composition may comprise a retinoic acid receptor agonist. A composition may comprise an agonist for the G protein-coupled receptor Smoothened. A composition may comprise a Dot1L inhibitor. A composition may comprise a Jak1/Jak2 inhibitor. A composition may comprise an SAH hydrolase inhibitor. A composition may comprise a c-Jun kinase inhibitor.

A composition may comprise a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor. A composition may comprise a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, and a serine-threonine kinase Akt inhibitor. A composition may comprise a ROCK inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, or a serine-threonine kinase Akt inhibitor, or any combination thereof. A composition may comprise a ROCK inhibitor, a c-Jun kinase inhibitor, or a CBP/p300 bromodomain inhibitor. A composition may comprise a ROCK inhibitor, a c-Jun kinase inhibitor, and a CBP/p300 bromodomain inhibitor.

A composition may comprise a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, and a c-Jun kinase inhibitor.

A composition may further comprise one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor. A composition may further comprise a G9a inhibitor or a DNMT inhibitor. A composition may further comprise an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or both.

A composition may comprise a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, a c-Jun kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, and a p38 MAPK inhibitor.

A composition may comprise somatic cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor, or any combination thereof. A composition may comprise somatic cells; and at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor. A composition may comprise somatic cells; and at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor.

A composition may comprise somatic cells; and a Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor. A composition may comprise somatic cells; and a CBP/p300 bromodomain inhibitor. A composition may comprise somatic cells; and a Menin-MLL interaction inhibitor. A composition may comprise a serine-threonine kinase Akt inhibitor. A composition may comprise somatic cells; and a glycogen kinase inhibitor. A composition may comprise somatic cells; and comprise a TGFβ receptor inhibitor. A composition may comprise somatic cells; and a retinoic acid receptor agonist. A composition may comprise somatic cells; and an agonist for the G protein-coupled receptor Smoothened. A composition may comprise somatic cells; and a Dot1L inhibitor. A composition may comprise somatic cells; and a Jak1/Jak2 inhibitor. A composition may comprise somatic cells; and an SAH hydrolase inhibitor. A composition may comprise somatic cells; and a c-Jun kinase inhibitor.

A composition may comprise somatic cells; and a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor. A composition may comprise somatic cells; and a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, and a serine-threonine kinase Akt inhibitor. A composition may comprise somatic cells; and a ROCK inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, or a serine-threonine kinase Akt inhibitor, or any combination thereof. A composition may comprise somatic cells; and a ROCK inhibitor, a c-Jun kinase inhibitor, or a CBP/p300 bromodomain inhibitor. A composition may comprise somatic cells; and a ROCK inhibitor, a c-Jun kinase inhibitor, and a CBP/p300 bromodomain inhibitor.

A composition may comprise somatic cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, and a c-Jun kinase inhibitor.

A composition comprising the somatic cells may further comprise one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor. A composition comprising the somatic cells may further comprise a G9a inhibitor or a DNMT inhibitor. A composition comprising the somatic cells may further comprise an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or both.

A composition may comprise somatic cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, a c-Jun kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, and a p38 MAPK inhibitor.

A composition may comprise intermediate plastic state cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor, or any combination thereof. A composition may comprise intermediate plastic state cells; and at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor. A composition may comprise intermediate plastic state cells; and at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 of a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, or a c-Jun kinase inhibitor.

A composition may comprise intermediate plastic state cells; and a Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor. A composition may comprise intermediate plastic state cells; and a CBP/p300 bromodomain inhibitor. A composition may comprise intermediate plastic state cells; and a Menin-MLL interaction inhibitor. A composition may comprise a serine-threonine kinase Akt inhibitor. A composition may comprise intermediate plastic state cells; and a glycogen kinase inhibitor. A composition may comprise intermediate plastic state cells; and comprise a TGFβ receptor inhibitor. A composition may comprise intermediate plastic state cells; and a retinoic acid receptor agonist. A composition may comprise intermediate plastic state cells; and an agonist for the G protein-coupled receptor Smoothened. A composition may comprise intermediate plastic state cells; and a Dot1L inhibitor. A composition may comprise intermediate plastic state cells; and a Jak1/Jak2 inhibitor. A composition may comprise intermediate plastic state cells; and an SAH hydrolase inhibitor. A composition may comprise intermediate plastic state cells; and a c-Jun kinase inhibitor.

A composition may comprise intermediate plastic state cells; and a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor. A composition may comprise intermediate plastic state cells; and a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, and a serine-threonine kinase Akt inhibitor. A composition may comprise intermediate plastic state cells; and a ROCK inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, or a serine-threonine kinase Akt inhibitor, or any combination thereof. A composition may comprise intermediate plastic state cells; and a ROCK inhibitor, a c-Jun kinase inhibitor, or a CBP/p300 bromodomain inhibitor. A composition may comprise intermediate plastic state cells; and a ROCK inhibitor, a c-Jun kinase inhibitor, and a CBP/p300 bromodomain inhibitor.

A composition may comprise intermediate plastic state cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, and a c-Jun kinase inhibitor.

A composition comprising the intermediate plastic state cells may further comprise one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor. A composition comprising the intermediate plastic state cells may further comprise a G9a inhibitor or a DNMT inhibitor. A composition comprising the intermediate plastic state cells may further comprise an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or both.

A composition may comprise intermediate plastic state cells; and a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, an SAH hydrolase inhibitor, a c-Jun kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, and a p38 MAPK inhibitor.

The intermediate plastic state cell may express LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2, or any combination thereof. The intermediate plastic state cell may express LIN28A. The intermediate plastic state cell may express SALL4. The intermediate plastic state cell may express MSX2. The intermediate plastic state cell may express NMYC. The intermediate plastic state cell may express WNT4. The intermediate plastic state cell may express FGF19. The intermediate plastic state cell may express TOP2A. The intermediate plastic state cell may express MSX1. The intermediate plastic state cell may express HOXB9. The intermediate plastic state cell may express WT1. The intermediate plastic state cell may express GATA2. The intermediate plastic state cell may express HMGA2. The intermediate plastic state cell may express LEF1. The intermediate plastic state cell may express FGF9. The intermediate plastic state cell may express HOXA9. The intermediate plastic state cell may express HOZXA1. The intermediate plastic state cell may express PTCH1. The intermediate plastic state cell may express HOXA5. The intermediate plastic state cell may express CCND2. The intermediate plastic state cell may express SDC1. The intermediate plastic state cell may express TBX3. The intermediate plastic state cell may express BMP4. The intermediate plastic state cell may express IGF2. The intermediate plastic state cell may express one or more of LIN28A, SALL4, MSX2, NMYC, WNT4, FGF19, TOP2A, MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

FIG. 4 depicts exemplary compositions comprising chemical reprogramming factors and optional cells in stage 1. In FIG. 4, "A" represents a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor, or any combination thereof; "B" represents a ROCK inhibitor, an Menin-MLL interaction inhibitor, or both, the composition may or may not have either of these two compounds in group B; "C" represents a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, a Jak1/Jak2 inhibitor, or a SAH hydrolase inhibitor, or any combination thereof, the composition may or may not have any of these compounds in group C; "D" represents an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor, or any combination thereof, the composition may or may not have any of these compounds in group D; "E" represents intermediate plastic state cells or somatic cells, or both, the composition may or may not have any of the cells in group E. For example, A1B1C1D1D2E2 represents a composition that includes a c-Jun kinase inhibitor, a ROCK inhibitor, a glycogen kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, and somatic cells.

A composition may comprise Y-27632 or thiazovivin. A composition may comprise SGC-CBP30, I-CBP112, GNE272, or GNE409. A composition may comprise VTP50469, MI3454, or WDR5-IN-4. A composition may comprise AKT Kinase Inhibitor. A composition may comprise CHIR99021 or CHIR98014. A composition may comprise E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334. A composition may comprise TTNPB, Ch55, or AM580. A composition may comprise SAG, Purmorphamine, Hh-Ag1.5, or human SHH. A composition may comprise EPZ004777 or EPZ5676. A composition may comprise Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib. A composition may comprise DZNep, NepA, Adox, or DZA. A composition may comprise JNKIN8, JNKIN7, JNKIN5, or JNKIN12. A composition may comprise 5-Iodotubercidin or ABT 702. A composition may comprise CX-4945, TPP 22, or Ellagic acid. A composition may comprise Unc0224, Unc0638, Unc0642 , or Bix01294. A composition may comprise 5-Azacytidine, Decitabine, RG108, or SGI-1027. A composition may comprise BIRB796, SB203580, or SB202190.

A composition may comprise: Y-27632 or thiazovivin; SGC-CBP30, I-CBP112, GNE272, or GNE409; VTP50469, MI3454, or WDR5-IN-4; AKT Kinase Inhibitor; CHIR99021 or CHIR98014; E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334; TTNPB, Ch55, or AM580; SAG, Purmorphamine, Hh-Ag1.5, or human SHH; EPZ004777 or EPZ5676; Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib; DZNep, NepA, Adox, or DZA; or JNKIN8, JNKIN7, JNKIN5, or JNKIN12. A composition may comprise; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; SGC-CBP30, I-CBP112, GNE272, or GNE409; or AKT Kinase Inhibitor. A composition may comprise; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; SGC-CBP30, I-CBP112, GNE272, or GNE409; and AKT Kinase Inhibitor. A composition may comprise: Y-27632 or thiazovivin; SGC-CBP30, I-CBP112, GNE272, or GNE409; VTP50469, MI3454, or WDR5-IN-4; or AKT Kinase Inhibitor. A composition may comprise: Y-27632 or thiazovivin; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; or SGC-CBP30, I-CBP112, GNE272, or GNE409. A composition may comprise: Y-27632 or thiazovivin; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; and SGC-CBP30, I-CBP112, GNE272, or GNE409. A composition may comprise: Y-27632 or thiazovivin; SGC-CBP30, I-CBP112, GNE272, or GNE409; VTP50469, MI3454, or WDR5-IN-4; AKT Kinase Inhibitor; CHIR99021 or CHIR98014; E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334; TTNPB, Ch55, or AM580; SAG, Purmorphamine, Hh-Ag1.5, or human SHH; EPZ004777 or EPZ5676; Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib; DZNep, NepA, Adox, or DZA; or JNKIN8, JNKIN7, JNKIN5, or JNKIN12.

A composition may further comprise: 5-Iodotubercidin or ABT 702; CX-4945, TPP 22, or Ellagic acid; Unc0224, Unc0638, Unc0642 , or Bix01294; 5-Azacytidine , Decitabine, RG108, or SGI-1027; or BIRB796, SB203580, or SB202190. Alternatively, a composition may further comprise: Unc0224, Unc0638, Unc0642 , or Bix01294; or 5-Azacytidine , Decitabine, RG108, or SGI-1027. Alternatively, a composition may further comprise: 5-Iodotubercidin or ABT 702; or CX-4945, TPP 22, or Ellagic acid.

A composition may comprise: Y-27632 or thiazovivin; SGC-CBP30, I-CBP112, GNE272, or GNE409; VTP50469, MI3454, or WDR5-IN-4; AKT Kinase Inhibitor; CHIR99021 or CHIR98014; E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334; TTNPB, Ch55, or AM580; SAG, Purmorphamine, Hh-Ag1.5, or human SHH; EPZ004777 or EPZ5676; Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib; DZNep, NepA, Adox, or DZA; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; 5-Iodotubercidin or ABT 702; CX-4945, TPP 22, or Ellagic acid; Unc0224, Unc0638, Unc0642 , or Bix01294; 5-Azacytidine , Decitabine, RG108, or SGI-1027; or BIRB796, SB203580, or SB202190.

A composition may comprise: Y-27632 or thiazovivin; SGC-CBP30, I-CBP112, GNE272, or GNE409; VTP50469, MI3454, or WDR5-IN-4; AKT Kinase Inhibitor; CHIR99021 or CHIR98014; E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334; TTNPB, Ch55, or AM580; SAG, Purmorphamine, Hh-Ag1.5, or human SHH; EPZ004777 or EPZ5676; Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib; DZNep, NepA, Adox, or DZA; JNKIN8, JNKIN7, JNKIN5, or JNKIN12; 5-Iodotubercidin or ABT 702; CX-4945, TPP 22, or Ellagic acid; Unc0224, Unc0638, Unc0642 , or Bix01294; 5-Azacytidine , Decitabine, RG108, or SGI-1027; and BIRB796, SB203580, or SB202190.

A composition may comprise Y-27632. A composition may comprise SGC-CBP30. A composition may comprise VTP50469. A composition may comprise AKT Kinase Inhibitor (AKTi) . A composition may comprise CHIR99021. A composition may comprise E-616452. A composition may comprise TTNPB. A composition may comprise SAG. A composition may comprise EPZ5676. A composition may comprise Ruxolitinib. A composition may comprise DZNep. A composition may comprise JNKIN8. A composition may comprise 5-Iodotubercidin. A composition may comprise CX-4945. A composition may comprise Unc0224. A composition may comprise 5-Azacytidine. A composition may comprise BIRB796.

A composition may comprise: Y-27632; SGC-CBP30; VTP50469; AKT Kinase Inhibitor; CHIR99021; E-616452; TTNPB; SAG; EPZ5676; Ruxolitinib; DZNep; or JNKIN8. A composition may comprise; JNKIN8; SGC-CBP30; or AKT Kinase Inhibitor. A composition may comprise; JNKIN8; SGC-CBP30; and AKT Kinase Inhibitor. A composition may comprise; Y-27632; SGC-CBP30; VTP50469; or AKT Kinase Inhibitor. A composition may comprise; Y-27632; JNKIN8; or SGC-CBP30. A composition may comprise; Y-27632; JNKIN8; and SGC-CBP30. A composition may comprise; Y-27632; SGC-CBP30; VTP50469; AKT Kinase Inhibitor; CHIR99021; E-616452; TTNPB; SAG; EPZ5676; Ruxolitinib; DZNep; or JNKIN8.

A composition may further comprise; 5-Iodotubercidin; CX-4945; Unc0224; 5-Azacytidine; or BIRB796. Alternatively, a composition may further comprise; Unc0224; or 5-Azacytidine. Alternatively, a composition may further comprise; 5-Iodotubercidin; or CX-4945.

A composition may comprise; Y-27632; SGC-CBP30; VTP50469; AKT Kinase Inhibitor; CHIR99021; E-616452; TTNPB; SAG; EPZ5676; Ruxolitinib; DZNep; JNKIN8; 5-Iodotubercidin; CX-4945; Unc0224; 5-Azacytidine; or BIRB796. A composition may comprise; Y-27632; SGC-CBP30; VTP50469; AKT Kinase Inhibitor; CHIR99021; E-616452; TTNPB; SAG; EPZ5676; Ruxolitinib; DZNep; JNKIN8; 5-Iodotubercidin; CX-4945; Unc0224; 5-Azacytidine; and BIRB796.

A composition may comprise at least about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, 500 μM or more Y-27632 within the composition. A composition may comprise at most about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, or 500 μM Y-27632 within the composition. A composition may comprise about 1 μM Y-27632 within the composition. A composition may comprise about 2 μM Y-27632 within the composition. A composition may comprise about 3 μM Y-27632 within the composition. A composition may comprise about 4 μM Y-27632 within the composition. A composition may comprise about 5 μM Y-27632 within the composition. A composition may comprise about 6 μM Y-27632 within the composition. A composition may comprise about 7 μM Y-27632 within the composition. A composition may comprise about 8 μM Y-27632 within the composition. A composition may comprise about 9 μM Y-27632 within the composition. A composition may comprise about 10 μM Y-27632 within the composition. A composition may comprise about 15 μM Y-27632 within the composition. A composition may comprise about 20 μM Y-27632 within the composition. A composition may comprise about 30 μM Y-27632 within the composition. A composition may comprise about 40 μM Y-27632 within the composition. A composition may comprise about 50 μM Y-27632 within the composition. A composition may comprise about 60 μM Y-27632 within the composition. A composition may comprise about 70 μM Y-27632 within the composition. A composition may comprise about 80 μM Y-27632 within the composition. A composition may comprise about 90 μM Y-27632 within the composition. A composition may comprise about 100 μM Y-27632 within the composition. A composition may comprise about 1 μM to about 100 μM Y-27632 within the composition. A composition may comprise about 2 μM to about 75 μM Y-27632 within the composition. A composition may comprise about 3 μM to about 50 μM Y-27632 within the composition. A composition may comprise about 4 μM to about 40 μM Y-27632 within the composition. A composition may comprise about 5 μM to about 30 μM Y-27632 within the composition. A composition may comprise about 7.5 μM to about 20 μM Y-27632 within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more SGC-CBP30 within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM SGC-CBP30 within the composition. A composition may comprise about 0.2 μM SGC-CBP30 within the composition. A composition may comprise about 0.4 μM SGC-CBP30 within the composition. A composition may comprise about 0.6 μM SGC-CBP30 within the composition. A composition may comprise about 0.8 μM SGC-CBP30 within the composition. A composition may comprise about 1 μM SGC-CBP30 within the composition. A composition may comprise about 1.2 μM SGC-CBP30 within the composition. A composition may comprise about 1.4 μM SGC-CBP30 within the composition. A composition may comprise about 1.6 μM SGC-CBP30 within the composition. A composition may comprise about 1.8 μM SGC-CBP30 within the composition. A composition may comprise about 2 μM SGC-CBP30 within the composition. A composition may comprise about 4 μM SGC-CBP30 within the composition. A composition may comprise about 6 μM SGC-CBP30 within the composition. A composition may comprise about 8 μM SGC-CBP30 within the composition. A composition may comprise about 10 μM SGC-CBP30 within the composition. A composition may comprise about 12 μM SGC-CBP30 within the composition. A composition may comprise about 14 μM SGC-CBP30 within the composition. A composition may comprise about 16 μM SGC-CBP30 within the composition. A composition may comprise about 18 μM SGC-CBP30 within the composition. A composition may comprise about 20 μM SGC-CBP30 within the composition. A composition may comprise about 0.2 μM to about 20 μM SGC-CBP30 within the composition. A composition may comprise about 0.4 μM to about 15 μM SGC-CBP30 within the composition. A composition may comprise about 0.6 μM to about 10 μM SGC-CBP30 within the composition. A composition may comprise about 0.8 μM to about 8 μM SGC-CBP30 within the composition. A composition may comprise about 1 μM to about 6 μM SGC-CBP30 within the composition. A composition may comprise about 1.5 μM to about 4 μM SGC-CBP30 within the composition.

A composition may comprise at least about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, 25 μM or more VTP50469 within the composition. A composition may comprise at most about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, or 25 μM VTP50469 within the composition. A composition may comprise about 0.05 μM VTP50469 within the composition. A composition may comprise about 0.1 μM VTP50469 within the composition. A composition may comprise about 0.15 μM VTP50469 within the composition. A composition may comprise about 0.2 μM VTP50469 within the composition. A composition may comprise about 0.25 μM VTP50469 within the composition. A composition may comprise about 0.3 μM VTP50469 within the composition. A composition may comprise about 0.35 μM VTP50469 within the composition. A composition may comprise about 0.4 μM VTP50469 within the composition. A composition may comprise about 0.45 μM VTP50469 within the composition. A composition may comprise about 0.5 μM VTP50469 within the composition. A composition may comprise about 1 μM VTP50469 within the composition. A composition may comprise about 1.5 μM VTP50469 within the composition. A composition may comprise about 2 μM VTP50469 within the composition. A composition may comprise about 2.5 μM VTP50469 within the composition. A composition may comprise about 3 μM VTP50469 within the composition. A composition may comprise about 3.5 μM VTP50469 within the composition. A composition may comprise about 4 μM VTP50469 within the composition. A composition may comprise about 4.5 μM VTP50469 within the composition. A composition may comprise about 5 μM VTP50469 within the composition. A composition may comprise about 0.05 μM to about 2.5 μM VTP50469 within the composition. A composition may comprise about 0.1 μM to about 1.875 μM VTP50469 within the composition. A composition may comprise about 0.15 μM to about 1.25 μM VTP50469 within the composition. A composition may comprise about 0.2 μM to about 1 μM VTP50469 within the composition. A composition may comprise about 0.25 μM to about 0.75 μM VTP50469 within the composition. A composition may comprise about 0.375 μM to about 0.5 μM VTP50469 within the composition.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, 50 μM or more AKTi within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, or 50 μM AKTi within the composition. A composition may comprise about 0.1 μM AKTi within the composition. A composition may comprise about 0.2 μM AKTi within the composition. A composition may comprise about 0.3 μM AKTi within the composition. A composition may comprise about 0.4 μM AKTi within the composition. A composition may comprise about 0.5 μM AKTi within the composition. A composition may comprise about 0.6 μM AKTi within the composition. A composition may comprise about 0.7 μM AKTi within the composition. A composition may comprise about 0.8 μM AKTi within the composition. A composition may comprise about 0.9 μM AKTi within the composition. A composition may comprise about 1 μM AKTi within the composition. A composition may comprise about 2 μM AKTi within the composition. A composition may comprise about 3 μM AKTi within the composition. A composition may comprise about 4 μM AKTi within the composition. A composition may comprise about 5 μM AKTi within the composition. A composition may comprise about 6 μM AKTi within the composition. A composition may comprise about 7 μM AKTi within the composition. A composition may comprise about 8 μM AKTi within the composition. A composition may comprise about 9 μM AKTi within the composition. A composition may comprise about 10 μM AKTi within the composition. A composition may comprise about 0.1 μM to about 10 μM AKTi within the composition. A composition may comprise about 0.2 μM to about 7.5 μM AKTi within the composition. A composition may comprise about 0.3 μM to about 5 μM AKTi within the composition. A composition may comprise about 0.4 μM to about 4 μM AKTi within the composition. A composition may comprise about 0.5 μM to about 3 μM AKTi within the composition. A composition may comprise about 0.75 μM to about 2 μM AKTi within the composition.

A composition may comprise at least about 0.1 micromolar (μM) , 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 10.5 μM, 11 μM, 11.5 μM, 12 μM, 12.5 μM, 13 μM, 13.5 μM, 14 μM, 14.5 μM, 15 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 26 μM, 27 μM, 28 μM, 29 μM, 30 μM, 31 μM, 32 μM, 33 μM, 34 μM, 35 μM, 36 μM, 37 μM, 38 μM, 39 μM, 40 μM, 41 μM, 42 μM, 43 μM, 44 μM, 45 μM, 46 μM, 47 μM, 48 μM, 49 μM, 50 μM, 100 μM, 150 μM, 200 μM, 250 μM or more CHIR99021 within the composition. A composition may comprise at most about 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 10.5 μM, 11 μM, 11.5 μM, 12 μM, 12.5 μM, 13 μM, 13.5 μM, 14 μM, 14.5 μM, 15 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 26 μM, 27 μM, 28 μM, 29 μM, 30 μM, 31 μM, 32 μM, 33 μM, 34 μM, 35 μM, 36 μM, 37 μM, 38 μM, 39 μM, 40 μM, 41 μM, 42 μM, 43 μM, 44 μM, 45 μM, 46 μM, 47 μM, 48 μM, 49 μM, 50 μM, 100 μM, 150 μM, 200 μM, or 250 μM CHIR99021 within the composition. A composition may comprise about 0.5 μM CHIR99021 within the composition. A composition may comprise about 1 μM CHIR99021 within the composition. A composition may comprise about 2 μM CHIR99021 within the composition. A composition may comprise about 3 μM CHIR99021 within the composition. A composition may comprise about 4 μM CHIR99021 within the composition. A composition may comprise about 5 μM CHIR99021 within the composition. A composition may comprise about 6 μM CHIR99021 within the composition. A composition may comprise about 7 μM CHIR99021 within the composition. A composition may comprise about 8 μM CHIR99021 within the composition. A composition may comprise about 9 μM CHIR99021 within the composition. A composition may comprise about 10 μM CHIR99021 within the composition. A composition may comprise about 15 μM CHIR99021 within the composition. A composition may comprise about 20 μM CHIR99021 within the composition. A composition may comprise about 30 μM CHIR99021 within the composition. A composition may comprise about 40 μM CHIR99021 within the composition. A composition may comprise about 50 μM CHIR99021 within the composition. A composition may comprise about 0.1 μM to about 100 μM CHIR99021 within the composition. A composition may comprise about 0.2 μM to about 75 μM CHIR99021 within the composition. A composition may comprise about 0.5 μM to about 50 μM CHIR99021 within the composition. A composition may comprise about 1 μM to about 25 μM CHIR99021 within the composition. A composition may comprise about 2 μM to about 12.5 μM CHIR99021 within the composition. A composition may comprise about 4 μM to about 6.25 μM CHIR99021 within the composition.

A composition may comprise at least about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, 500 μM or more E-616452 within the composition. A composition may comprise at most about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, or 500 μM E-616452 within the composition. A composition may comprise about 1 μM E-616452 within the composition. A composition may comprise about 2 μM E-616452 within the composition. A composition may comprise about 3 μM E-616452 within the composition. A composition may comprise about 4 μM E-616452 within the composition. A composition may comprise about 5 μM E-616452 within the composition. A composition may comprise about 6 μM E-616452 within the composition. A composition may comprise about 7 μM E-616452 within the composition. A composition may comprise about 8 μM E-616452 within the composition. A composition may comprise about 9 μM E-616452 within the composition. A composition may comprise about 10 μM E-616452 within the composition. A composition may comprise about 15 μM E-616452 within the composition. A composition may comprise about 20 μM E-616452 within the composition. A composition may comprise about 30 μM E-616452 within the composition. A composition may comprise about 40 μM E-616452 within the composition. A composition may comprise about 50 μM E-616452 within the composition. A composition may comprise about 60 μM E-616452 within the composition. A composition may comprise about 70 μM E-616452 within the composition. A composition may comprise about 80 μM E-616452 within the composition. A composition may comprise about 90 μM E-616452 within the composition. A composition may comprise about 100 μM E-616452 within the composition. A composition may comprise about 1 μM to about 100 μM E-616452 within the composition. A composition may comprise about 2 μM to about 75 μM E-616452 within the composition. A composition may comprise about 3 μM to about 50 μM E-616452 within the composition. A composition may comprise about 4 μM to about 40 μM E-616452 within the composition. A composition may comprise about 5 μM to about 30 μM E-616452 within the composition. A composition may comprise about 7.5 μM to about 20 μM E-616452 within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more TTNPB within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM TTNPB within the composition. A composition may comprise about 0.2 μM TTNPB within the composition. A composition may comprise about 0.4 μM TTNPB within the composition. A composition may comprise about 0.6 μM TTNPB within the composition. A composition may comprise about 0.8 μM TTNPB within the composition. A composition may comprise about 1 μM TTNPB within the composition. A composition may comprise about 1.2 μM TTNPB within the composition. A composition may comprise about 1.4 μM TTNPB within the composition. A composition may comprise about 1.6 μM TTNPB within the composition. A composition may comprise about 1.8 μM TTNPB within the composition. A composition may comprise about 2 μM TTNPB within the composition. A composition may comprise about 4 μM TTNPB within the composition. A composition may comprise about 6 μM TTNPB within the composition. A composition may comprise about 8 μM TTNPB within the composition. A composition may comprise about 10 μM TTNPB within the composition. A composition may comprise about 12 μM TTNPB within the composition. A composition may comprise about 14 μM TTNPB within the composition. A composition may comprise about 16 μM TTNPB within the composition. A composition may comprise about 18 μM TTNPB within the composition. A composition may comprise about 20 μM TTNPB within the composition. A composition may comprise about 0.2 μM to about 20 μM TTNPB within the composition. A composition may comprise about 0.4 μM to about 15 μM TTNPB within the composition. A composition may comprise about 0.6 μM to about 10 μM TTNPB within the composition. A composition may comprise about 0.8 μM to about 8 μM TTNPB within the composition. A composition may comprise about 1 μM to about 6 μM TTNPB within the composition. A composition may comprise about 1.5 μM to about 4 μM TTNPB within the composition.

A composition may comprise at least about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, 25 μM or more SAG within the composition. A composition may comprise at most about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, or 25 μM SAG within the composition. A composition may comprise about 0.05 μM SAG within the composition. A composition may comprise about 0.1 μM SAG within the composition. A composition may comprise about 0.15 μM SAG within the composition. A composition may comprise about 0.2 μM SAG within the composition. A composition may comprise about 0.25 μM SAG within the composition. A composition may comprise about 0.3 μM SAG within the composition. A composition may comprise about 0.35 μM SAG within the composition. A composition may comprise about 0.4 μM SAG within the composition. A composition may comprise about 0.45 μM SAG within the composition. A composition may comprise about 0.5 μM SAG within the composition. A composition may comprise about 1 μM SAG within the composition. A composition may comprise about 1.5 μM SAG within the composition. A composition may comprise about 2 μM SAG within the composition. A composition may comprise about 2.5 μM SAG within the composition. A composition may comprise about 3 μM SAG within the composition. A composition may comprise about 3.5 μM SAG within the composition. A composition may comprise about 4 μM SAG within the composition. A composition may comprise about 4.5 μM SAG within the composition. A composition may comprise about 5 μM SAG within the composition. A composition may comprise about 0.05 μM to about 2.5 μM SAG within the composition. A composition may comprise about 0.1 μM to about 1.875 μM SAG within the composition. A composition may comprise about 0.15 μM to about 1.25 μM SAG within the composition. A composition may comprise about 0.2 μM to about 1 μM SAG within the composition. A composition may comprise about 0.25 μM to about 0.75 μM SAG within the composition. A composition may comprise about 0.375 μM to about 0.5 μM SAG within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more EPZ5676 within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM EPZ5676 within the composition. A composition may comprise about 0.2 μM EPZ5676 within the composition. A composition may comprise about 0.4 μM EPZ5676 within the composition. A composition may comprise about 0.6 μM EPZ5676 within the composition. A composition may comprise about 0.8 μM EPZ5676 within the composition. A composition may comprise about 1 μM EPZ5676 within the composition. A composition may comprise about 1.2 μM EPZ5676 within the composition. A composition may comprise about 1.4 μM EPZ5676 within the composition. A composition may comprise about 1.6 μM EPZ5676 within the composition. A composition may comprise about 1.8 μM EPZ5676 within the composition. A composition may comprise about 2 μM EPZ5676 within the composition. A composition may comprise about 3 μM EPZ5676 within the composition. A composition may comprise about 4 μM EPZ5676 within the composition. A composition may comprise about 6 μM EPZ5676 within the composition. A composition may comprise about 8 μM EPZ5676 within the composition. A composition may comprise about 10 μM EPZ5676 within the composition. A composition may comprise about 12 μM EPZ5676 within the composition. A composition may comprise about 14 μM EPZ5676 within the composition. A composition may comprise about 16 μM EPZ5676 within the composition. A composition may comprise about 18 μM EPZ5676 within the composition. A composition may comprise about 20 μM EPZ5676 within the composition. A composition may comprise about 0.2 μM to about 20 μM EPZ5676 within the composition. A composition may comprise about 0.4 μM to about 15 μM EPZ5676 within the composition. A composition may comprise about 0.6 μM to about 10 μM EPZ5676 within the composition. A composition may comprise about 0.8 μM to about 8 μM EPZ5676 within the composition. A composition may comprise about 1 μM to about 6 μM EPZ5676 within the composition. A composition may comprise about 1.5 μM to about 4 μM EPZ5676 within the composition.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, 50 μM or more Ruxolitinib within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, or 50 μM Ruxolitinib within the composition. A composition may comprise about 0.1 μM Ruxolitinib within the composition. A composition may comprise about 0.2 μM Ruxolitinib within the composition. A composition may comprise about 0.3 μM Ruxolitinib within the composition. A composition may comprise about 0.4 μM Ruxolitinib within the composition. A composition may comprise about 0.5 μM Ruxolitinib within the composition. A composition may comprise about 0.6 μM Ruxolitinib within the composition. A composition may comprise about 0.7 μM Ruxolitinib within the composition. A composition may comprise about 0.8 μM Ruxolitinib within the composition. A composition may comprise about 0.9 μM Ruxolitinib within the composition. A composition may comprise about 1 μM Ruxolitinib within the composition. A composition may comprise about 2 μM Ruxolitinib within the composition. A composition may comprise about 3 μM Ruxolitinib within the composition. A composition may comprise about 4 μM Ruxolitinib within the composition. A composition may comprise about 5 μM Ruxolitinib within the composition. A composition may comprise about 6 μM Ruxolitinib within the composition. A composition may comprise about 7 μM Ruxolitinib within the composition. A composition may comprise about 8 μM Ruxolitinib within the composition. A composition may comprise about 9 μM Ruxolitinib within the composition. A composition may comprise about 10 μM Ruxolitinib within the composition. A composition may comprise about 0.1 μM to about 10 μM Ruxolitinib within the composition. A composition may comprise about 0.2 μM to about 7.5 μM Ruxolitinib within the composition. A composition may comprise about 0.3 μM to about 5 μM Ruxolitinib within the composition. A composition may comprise about 0.4 μM to about 4 μM Ruxolitinib within the composition. A composition may comprise about 0.5 μM to about 3 μM Ruxolitinib within the composition. A composition may comprise about 0.75 μM to about 2 μM Ruxolitinib within the composition.

A composition may comprise at least about 0.0004 μM, 0.0008 μM, 0.0012 μM, 0.0016 μM, 0.002 μM, 0.004 μM, 0.006 μM, 0.008 μM, 0.01 μM, 0.012 μM, 0.014 μM, 0.016 μM, 0.018 μM, 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1 μM or more DZNep within the composition. A composition may comprise at most about 0.0004 μM, 0.0008 μM, 0.0012 μM, 0.0016 μM, 0.002 μM, 0.004 μM, 0.006 μM, 0.008 μM, 0.01 μM, 0.012 μM, 0.014 μM, 0.016 μM, 0.018 μM, 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, or 1 μM DZNep within the composition. A composition may comprise about 0.01 μM DZNep within the composition. A composition may comprise about 0.012 μM DZNep within the composition. A composition may comprise about 0.014 μM DZNep within the composition. A composition may comprise about 0.016 μM DZNep within the composition. A composition may comprise about 0.018 μM DZNep within the composition. A composition may comprise about 0.02 μM DZNep within the composition. A composition may comprise about 0.04 μM DZNep within the composition. A composition may comprise about 0.06 μM DZNep within the composition. A composition may comprise about 0.08 μM DZNep within the composition. A composition may comprise about 0.1 μM DZNep within the composition. A composition may comprise about 0.12 μM DZNep within the composition. A composition may comprise about 0.14 μM DZNep within the composition. A composition may comprise about 0.16 μM DZNep within the composition. A composition may comprise about 0.18 μM DZNep within the composition. A composition may comprise about 0.2 μM DZNep within the composition. A composition may comprise about 0.22 μM DZNep within the composition. A composition may comprise about 0.24 μM DZNep within the composition. A composition may comprise about 0.26 μM DZNep within the composition. A composition may comprise about 0.28 μM DZNep within the composition. A composition may comprise about 0.3 μM DZNep within the composition. A composition may comprise about 0.4 μM DZNep within the composition. A composition may comprise about 0.5 μM DZNep within the composition. A composition may comprise about 0.6 μM DZNep within the composition. A composition may comprise about 0.7 μM DZNep within the composition. A composition may comprise about 0.8 μM DZNep within the composition. A composition may comprise about 0.9 μM DZNep within the composition. A composition may comprise about 1.0 μM DZNep within the composition. A composition may comprise about 0.02 μM to about 2 μM DZNep within the composition. A composition may comprise about 0.025 μM to about 1.5 μM DZNep within the composition. A composition may comprise about 0.05 μM to about 1 μM DZNep within the composition. A composition may comprise about 0.075 μM to about 0.75 μM DZNep within the composition. A composition may comprise about 0.1 μM to about 0.5 μM DZNep within the composition. A composition may comprise about 0.15 μM to about 0.4 μM DZNep within the composition.

A composition may comprise at least about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, 25 μM or more JNKIN8 within the composition. A composition may comprise at most about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, or 25 μM JNKIN8 within the composition. A composition may comprise about 0.05 μM JNKIN8 within the composition. A composition may comprise about 0.1 μM JNKIN8 within the composition. A composition may comprise about 0.15 μM JNKIN8 within the composition. A composition may comprise about 0.2 μM JNKIN8 within the composition. A composition may comprise about 0.25 μM JNKIN8 within the composition. A composition may comprise about 0.3 μM JNKIN8 within the composition. A composition may comprise about 0.35 μM JNKIN8 within the composition. A composition may comprise about 0.4 μM JNKIN8 within the composition. A composition may comprise about 0.45 μM JNKIN8 within the composition. A composition may comprise about 0.5 μM JNKIN8 within the composition. A composition may comprise about 1 μM JNKIN8 within the composition. A composition may comprise about 1.5 μM JNKIN8 within the composition. A composition may comprise about 2 μM JNKIN8 within the composition. A composition may comprise about 2.5 μM JNKIN8 within the composition. A composition may comprise about 3 μM JNKIN8 within the composition. A composition may comprise about 3.5 μM JNKIN8 within the composition. A composition may comprise about 4 μM JNKIN8 within the composition. A composition may comprise about 4.5 μM JNKIN8 within the composition. A composition may comprise about 5 μM JNKIN8 within the composition. A composition may comprise about 0.05 μM to about 2.5 μM JNKIN8 within the composition. A composition may comprise about 0.1 μM to about 1.875 μM JNKIN8 within the composition. A composition may comprise about 0.15 μM to about 1.25 μM JNKIN8 within the composition. A composition may comprise about 0.2 μM to about 1 μM JNKIN8 within the composition. A composition may comprise about 0.25 μM to about 0.75 μM JNKIN8 within the composition. A composition may comprise about 0.375 μM to about 0.5 μM JNKIN8 within the composition.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, 50 μM or more 5-Iodotubercidin within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, or 50 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.1 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.2 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.3 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.4 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.5 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.6 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.7 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.8 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.9 μM 5-Iodotubercidin within the composition. A composition may comprise about 1 μM 5-Iodotubercidin within the composition. A composition may comprise about 2 μM 5-Iodotubercidin within the composition. A composition may comprise about 3 μM 5-Iodotubercidin within the composition. A composition may comprise about 4 μM 5-Iodotubercidin within the composition. A composition may comprise about 5 μM 5-Iodotubercidin within the composition. A composition may comprise about 6 μM 5-Iodotubercidin within the composition. A composition may comprise about 7 μM 5-Iodotubercidin within the composition. A composition may comprise about 8 μM 5-Iodotubercidin within the composition. A composition may comprise about 9 μM 5-Iodotubercidin within the composition. A composition may comprise about 10 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.1 μM to about 10 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.2 μM to about 7.5 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.3 μM to about 5 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.4 μM to about 4 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.5 μM to about 3 μM 5-Iodotubercidin within the composition. A composition may comprise about 0.75 μM to about 2 μM 5-Iodotubercidin within the composition.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, 50 μM or more unc022 within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, or 50 μM unc022 within the composition. A composition may comprise about 0.1 μM unc022 within the composition. A composition may comprise about 0.2 μM unc022 within the composition. A composition may comprise about 0.3 μM unc022 within the composition. A composition may comprise about 0.4 μM unc022 within the composition. A composition may comprise about 0.5 μM unc022 within the composition. A composition may comprise about 0.6 μM unc022 within the composition. A composition may comprise about 0.7 μM unc022 within the composition. A composition may comprise about 0.8 μM unc022 within the composition. A composition may comprise about 0.9 μM unc022 within the composition. A composition may comprise about 1 μM unc022 within the composition. A composition may comprise about 2 μM unc022 within the composition. A composition may comprise about 3 μM unc022 within the composition. A composition may comprise about 4 μM unc022 within the composition. A composition may comprise about 5 μM unc022 within the composition. A composition may comprise about 6 μM unc022 within the composition. A composition may comprise about 7 μM unc022 within the composition. A composition may comprise about 8 μM unc022 within the composition. A composition may comprise about 9 μM unc022 within the composition. A composition may comprise about 10 μM unc022 within the composition. A composition may comprise about 0.1 μM to about 10 μM unc022 within the composition. A composition may comprise about 0.2 μM to about 7.5 μM unc022 within the composition. A composition may comprise about 0.3 μM to about 5 μM unc022 within the composition. A composition may comprise about 0.4 μM to about 4 μM unc022 within the composition. A composition may comprise about 0.5 μM to about 3 μM unc022 within the composition. A composition may comprise about 0.75 μM to about 2 μM unc022 within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more 5-Azacytidine within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM 5-Azacytidine within the composition. A composition may comprise about 0.2 μM 5-Azacytidine within the composition. A composition may comprise about 0.4 μM 5-Azacytidine within the composition. A composition may comprise about 0.6 μM 5-Azacytidine within the composition. A composition may comprise about 0.8 μM 5-Azacytidine within the composition. A composition may comprise about 1 μM 5-Azacytidine within the composition. A composition may comprise about 1.2 μM 5-Azacytidine within the composition. A composition may comprise about 1.4 μM 5-Azacytidine within the composition. A composition may comprise about 1.6 μM 5-Azacytidine within the composition. A composition may comprise about 1.8 μM 5-Azacytidine within the composition. A composition may comprise about 2 μM 5-Azacytidine within the composition. A composition may comprise about 4 μM 5-Azacytidine within the composition. A composition may comprise about 6 μM 5-Azacytidine within the composition. A composition may comprise about 8 μM 5-Azacytidine within the composition. A composition may comprise about 10 μM 5-Azacytidine within the composition. A composition may comprise about 12 μM 5-Azacytidine within the composition. A composition may comprise about 14 μM 5-Azacytidine within the composition. A composition may comprise about 16 μM 5-Azacytidine within the composition. A composition may comprise about 18 μM 5-Azacytidine within the composition. A composition may comprise about 20 μM 5-Azacytidine within the composition. A composition may comprise about 0.2 μM to about 20 μM 5-Azacytidine within the composition. A composition may comprise about 0.4 μM to about 15 μM 5-Azacytidine within the composition. A composition may comprise about 0.6 μM to about 10 μM 5-Azacytidine within the composition. A composition may comprise about 0.8 μM to about 8 μM 5-Azacytidine within the composition. A composition may comprise about 1 μM to about 6 μM 5-Azacytidine within the composition. A composition may comprise about 1.5 μM to about 4 μM 5-Azacytidine within the composition.

A composition may comprise at least about 0.016 μM, 0.018 μM, 0.02 μM, 0.022 μM, 0.024 μM, 0.026 μM, 0.028 μM, 0.03 μM, 0.032 μM, 0.034 μM, 0.036 μM, 0.038 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.09 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM or more CX-4945 within the composition. A composition may comprise at most about 0.016 μM, 0.018 μM, 0.02 μM, 0.022 μM, 0.024 μM, 0.026 μM, 0.028 μM, 0.03 μM, 0.032 μM, 0.034 μM, 0.036 μM, 0.038 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.09 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM CX-4945 within the composition. A composition may comprise about 0.08 μM CX-4945 within the composition. A composition may comprise about 0.1 μM CX-4945 within the composition. A composition may comprise about 0.2 μM CX-4945 within the composition. A composition may comprise about 0.3 μM CX-4945 within the composition. A composition may comprise about 0.4 μM CX-4945 within the composition. A composition may comprise about 0.5 μM CX-4945 within the composition. A composition may comprise about 0.6 μM CX-4945 within the composition. A composition may comprise about 0.7 μM CX-4945 within the composition. A composition may comprise about 0.8 μM CX-4945 within the composition. A composition may comprise about 0.9 μM CX-4945 within the composition. A composition may comprise about 1 μM CX-4945 within the composition. A composition may comprise about 1.5 μM CX-4945 within the composition. A composition may comprise about 2 μM CX-4945 within the composition. A composition may comprise about 3 μM CX-4945 within the composition. A composition may comprise about 4 μM CX-4945 within the composition. A composition may comprise about 5 μM CX-4945 within the composition. A composition may comprise about 6 μM CX-4945 within the composition. A composition may comprise about 7 μM CX-4945 within the composition. A composition may comprise about 8 μM CX-4945 within the composition. A composition may comprise about 0.08 μM to about 8 μM CX-4945 within the composition. A composition may comprise about 0.1 μM to about 4 μM CX-4945 within the composition. A composition may comprise about 0.15 μM to about 2 μM CX-4945 within the composition. A composition may comprise about 0.25 μM to about 1.5 μM CX-4945 within the composition. A composition may comprise about 0.5 μM to about 1 μM CX-4945 within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more BIRB796 within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM BIRB796 within the composition. A composition may comprise about 0.2 μM BIRB796 within the composition. A composition may comprise about 0.4 μM BIRB796 within the composition. A composition may comprise about 0.6 μM BIRB796 within the composition. A composition may comprise about 0.8 μM BIRB796 within the composition. A composition may comprise about 1 μM BIRB796 within the composition. A composition may comprise about 1.2 μM BIRB796 within the composition. A composition may comprise about 1.4 μM BIRB796 within the composition. A composition may comprise about 1.6 μM BIRB796 within the composition. A composition may comprise about 1.8 μM BIRB796 within the composition. A composition may comprise about 2 μM BIRB796 within the composition. A composition may comprise about 4 μM BIRB796 within the composition. A composition may comprise about 6 μM BIRB796 within the composition. A composition may comprise about 8 μM BIRB796 within the composition. A composition may comprise about 10 μM BIRB796 within the composition. A composition may comprise about 12 μM BIRB796 within the composition. A composition may comprise about 14 μM BIRB796 within the composition. A composition may comprise about 16 μM BIRB796 within the composition. A composition may comprise about 18 μM BIRB796 within the composition. A composition may comprise about 20 μM BIRB796 within the composition. A composition may comprise about 0.2 μM to about 20 μM BIRB796 within the composition. A composition may comprise about 0.4 μM to about 15 μM BIRB796 within the composition. A composition may comprise about 0.6 μM to about 10 μM BIRB796 within the composition. A composition may comprise about 0.8 μM to about 8 μM BIRB796 within the composition. A composition may comprise about 1 μM to about 6 μM BIRB796 within the composition. A composition may comprise about 1.5 μM to about 4 μM BIRB796 within the composition.

Subsequent to contacting any populations of stage 1 cells with any compositions described herein, the cells may be incubated in hypoxic condition. For example, the stage 1 cells may be incubated with at most 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%or lower atmospheric oxygen. The hypoxic condition may comprise about 10%atmospheric oxygen. The hypoxic condition may comprise about 9%atmospheric oxygen. The hypoxic condition may comprise about 8%atmospheric oxygen. The hypoxic condition may comprise about 7%atmospheric oxygen. The hypoxic condition may comprise about 6%atmospheric oxygen. The hypoxic condition may comprise about 5%atmospheric oxygen. The hypoxic condition may comprise about 4%atmospheric oxygen. The hypoxic condition may comprise about 3%atmospheric oxygen. The hypoxic condition may comprise about 2%atmospheric oxygen. The hypoxic condition may comprise about 1%atmospheric oxygen.

Subsequent to contacting any populations of stage 1 cells with any composition described herein, a population of stage 1 cells may be incubated with a composition for at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 25 days, or 30 days. A population of stage 1 cells may be incubated with a composition for at most about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 25 days, or 30 days. A population of stage 1 cells may be incubated with a composition for about 4 days. A population of stage 1 cells may be incubated with a composition for about 5 days. A population of stage 1 cells may be incubated with a composition for about 6 days. A population of stage 1 cells may be incubated with a composition for about 7 days. A population of stage 1 cells may be incubated with a composition for about 8 days. A population of stage 1 cells may be incubated with a composition for about 9 days. A population of stage 1 cells may be incubated with a composition for about 10 days. A population of stage 1 cells may be incubated with a composition for about 11 days. A population of stage 1 cells may be incubated with a composition for about 12 days. A population of stage 1 cells may be incubated with a composition for about 13 days. A population of stage 1 cells may be incubated with a composition for about 14 days. A population of stage 1 cells may be incubated with a composition for about 15 days. A population of stage 1 cells may be incubated with a composition for about 16 days. A population of stage 1 cells may be incubated with a composition for about 17 days. A population of stage 1 cells may be incubated with a composition for about 18 days. A population of stage 1 cells may be incubated with a composition for about 19 days. A population of stage 1 cells may be incubated with a composition for about 20 days. A population of stage 1 cells may be incubated with a composition for about 21 days. A population of stage 1 cells may be incubated with a composition for about 22 days. A population of stage 1 cells may be incubated with a composition for about 23 days. A population of stage 1 cells may be incubated with a composition for about 24 days. A population of stage 1 cells may be incubated with a composition for about 25 days. A population of stage 1 cells may be incubated with a composition for about 28 days. A population of stage 1 cells may be incubated with a composition for about 30 days.

Any of the compositions may not comprise feeder cells or serum. Any of the compositions may not comprise feeder cells and serum. Any of the compositions may not comprise feeder cells. Any of the compositions may be serum-free. Any of the compositions may comprise feeder cells. Any of the compositions may comprise serum.

STAGE 2

In some aspects, provided herein are stage 2 methods and compositions for conversion of an intermediate plastic state cell into a cell with a higher cell potency (e.g., less specialized cell) , such as a pluripotent stem cell –the conversion process referred herein also as "stage 2. " A stage 2 method may be part of a conversion process that reprograms intermediate plastic state cells into pluripotent stem cells. A stage 2 method may be the third stage of a conversion process that reprograms somatic plastic state cells into pluripotent stem cells.

A stage 2 method may comprise contacting a first cell population with a first composition. A stage 2 method may comprise, subsequent to or during the contacting, converting a subset of the first cell population into different cells. The cell population comprising the different cells may comprise a second cell population. A stage 2 method may comprise incubating the first cell population with the first composition for a period of time. The subset of the first cell population may be converted into the different cells prior to, during, or subsequent to the incubating. In some cases, a stage 2 method may comprise removing the first composition from the second population of cells. In other cases, a stage 2 method may comprise removing the first composition from the first population of cells.

A first population of stage 2 cells may comprise any populations of stage 1 cells. In some case, the first population of stage 2 cells may comprise the second population of stage 1 cells. In some case, the first population of stage 2 cells may comprise a composition in which the second population of stage 1 cells are isolated or removed from the stage 1 chemical reprogramming factors.

The second population of stage 2 cells may comprise pluripotent stem cells. the pluripotent stem cells obtained after contacting a population of cells with a composition may be referred to chemically induced pluripotent stem cells (CiPSCs) . CiPSCs may comprise human CiPSCs (hCiPSCs) . The first population of stage 2 cells may comprise pluripotent stem cells.

A CiPSC is not a naturally occurring cell. A CiPSC may express a combination of genes that are not expressed by a naturally occurring cell. In some cases, a CiPSC may express at least one gene at level at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 5-fold, 100-fold, or higher, relative to a naturally occurring cell. In some cases, a CiPSC may express at least one gene at level at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%lower, relative to a naturally occurring cell. The second population of stage 2 cells may comprise somatic cells, intermediate plastic state cells or CiPSCs. The second population of stage 2 cells may comprise somatic cells, intermediate plastic state cells and CiPSCs. The second population of stage 2 cells may not comprise somatic cells or intermediate plastic state cells. In some cases, the second population of stage 2 cells may comprise fewer somatic cells or intermediate plastic state cells than the first population of stage 2 cells. For example, the second population of stage 2 cells may have 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%or 99%fewer somatic cells or intermediate plastic state cells than the first population of stage 2 cells. In some cases, the second population of stage 2 cells may comprise more CiPSCs than the first population of stage 2 cells. For example, the second population of stage 2 cells may have 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 5-fold, 100-fold, or more CiPSCs than the first population of stage 2 cells.

A CiPSC may express OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, or any combination thereof. A CiPSC may express OCT4. A CiPSC may express SOX2. A CiPSC may express NANOG. A CiPSC may express FGF4. A CiPSC may express ZFP57. A CiPSC may express DPPA5. A CiPSC may express REX1. A CiPSC may express DPPA4. A CiPSC may express TDGF1. A CiPSC may express TRA-1-60. A CiPSC may express TRA-1-81. A CiPSC may express SSEA4. A CiPSC may express KLF4. A CiPSC may express KLF17. A CiPSC may express DPPA3. A CiPSC may express DNMT3L. A CiPSC may express UTF1. A CiPSC may express one or more of OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1.

A CiPSC may express OCT4, SOX2, or NANOG. A CiPSC may express OCT4, SOX2, and NANOG. A CiPSC may express OCT4 or SOX2. A CiPSC may express OCT4 or NANOG. A CiPSC may express SOX2 or NANOG. A CiPSC may express OCT4 and SOX2. A CiPSC may express OCT4 and NANOG. A CiPSC may express SOX2 and NANOG. A CiPSC may express OCT4, SOX2, and NANOG; and FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, or any combination thereof. A CiPSC may express OCT4, SOX2, and NANOG; and FGF4. A CiPSC may express OCT4, SOX2, and NANOG; and ZFP57. A CiPSC may express OCT4, SOX2, and NANOG; and DPPA5. A CiPSC may express OCT4, SOX2, and NANOG; and REX1. A CiPSC may express OCT4, SOX2, and NANOG; and DPPA4. A CiPSC may express OCT4, SOX2, and NANOG; and TDGF1. A CiPSC may express OCT4, SOX2, and NANOG; and TRA-1-60. A CiPSC may express OCT4, SOX2, and NANOG; and TRA-1-81. A CiPSC may express OCT4, SOX2, and NANOG; and SSEA4. A CiPSC may express OCT4, SOX2, and NANOG; and KLF4. A CiPSC may express OCT4, SOX2, and NANOG; and KLF17. A CiPSC may express OCT4, SOX2, and NANOG; and DPPA3. A CiPSC may express OCT4, SOX2, and NANOG; and DNMT3L. A CiPSC may express OCT4, SOX2, and NANOG; and UTF1. A CiPSC may express OCT4, SOX2, and NANOG; and one or more of FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DPPA5, DNMT3L, REX1, or UTF1.

A CPISC may express OCT4, SOX2, and NANOG; a second gene; and a third gene. The second gene expressed by the CPISC may comprise FGF4, ZFP57, DPPA5, or REX1, or any combination thereof. A somatic cell or an intermediate plastic state cell may not express OCT4. A somatic cell or an intermediate plastic state cell may not express SOX2. A somatic cell or an intermediate plastic state cell may not express NANOG. A somatic cell or an intermediate plastic state cell may not express OCT4, SOX2, or NANOG. A somatic cell or an intermediate plastic state cell may not express OCT4, SOX2, and NANOG. A somatic cell or an intermediate plastic state cell may not express OCT4, SOX2, or NANOG; a second gene; or a third gene. A somatic cell or an intermediate plastic state cell may not express the second gene. A somatic cell or an intermediate plastic state cell may not express the third gene. A somatic cell or an intermediate plastic state cell may not express OCT4, SOX2, or NANOG; one or more second genes; and one or more third genes.

The second gene may comprise FGF4, ZFP57, DPPA5, or REX1, or any combination thereof. The second gene may comprise FGF4. The second gene may comprise ZFP57. The second gene may comprise DPPA5. The second gene may comprise REX1. The third gene may comprise DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, or any combination thereof. The third gene may comprise DPPA4. The third gene may comprise TDGF1. The third gene may comprise TRA-1-60. The third gene may comprise TRA-1-81. The third gene may comprise SSEA4. The third gene may comprise KLF4. The third gene may comprise KLF17. The third gene may comprise DPPA3. The third gene may comprise DNMT3L. The third gene may comprise UTF1. FIG. 5 depicts exemplary CiPSCs that express OCT4, SOX2, and NANOG shown in a heatmap. The x-axis and y-axis of the heatmap show the second and the third genes, respectively. Each pixel of the heatmap represents one cell population. For example, cells 241 express OCT4, SOX2, NANOG, ZFP57, and SSEA4. Cells 242 express OCT4, SOX2, NANOG, REX1, or any combination of the third genes (e.g., TRA-1-60 and TRA-1-81) . Cells 243 express OCT4, SOX2, NANOG, KLF17, or any combination of the second genes (e.g., DNMT3L and UTF1) .

A cell of the second population of stage 2 cells may express higher levels of OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, or any combination thereof, relative to a cell of the first population of stage 2 cells or any populations of the stage 1 cells. The higher level of expression of any one of OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, or any combination thereof in a cell of the second population of stage 2 cells may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 5-fold, 100-fold, or more, relative to a cell of the first population of stage 2 cells or any populations of the stage 1 cells. A cell of the first population of stage 2 cells or any populations of the stage 1 cells may express lower levels of any one of OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1, relative to a cell of the second population of stage 2 cells. The lower level of expression of any one of OCT4, SOX2, NANOG, FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1 in a cell of the first population of stage 2 cells or any populations of the stage 1 cells may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%, relative to a cell of the second population of stage 2 cells. The levels of expression can be measured by any methods described herein. For examples, gene expression can be measured by methods described in EXAMPLE 2. Gene expression can be measured by using any one of SEQ ID NO: 1-83 (including controls) .

In some aspects, provided herein is a composition that comprises reprogramming factors for stage 2 conversion, or comprises cells of stage 2 (the first population of cells or the second population of cells) , or comprises cells of stage 2 (the first population of cells or the second population of cells) and reprogramming factors for stage 2 conversion. In some cases, a composition comprises a culture medium comprising the reprogramming factors for stage 2 conversion. In some cases, the composition is a culture medium comprising one or more chemical reprogramming factors provided herein.

In some cases, a composition may comprise an isolated population of the second population of stage 2 cells. In some cases, a composition may comprise an isolated population the first population of stage 2 cells. An isolated population of stage 2 cells may comprise at least about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more cells. An isolated population of stage 2 cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 cells. An isolated population of stage 2 cells may comprise at least one CiPSC. In some cases, an isolated population of stage 2 cells may comprise at least about 1, 1 x 10^1, 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more CiPSCs. An isolated population of cell may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 CiPSCs. In some cases, an isolated population of stage 2 cells may comprise at least about 1 x 10^1, 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, 1x10^10 or more CiPSCs, intermediate plastic state cells, or somatic cells, or any combination thereof. An isolated population of stage 2 cells may comprise at most about 1x10^2, 1x10^3, 1x10^4, 1x10^5, 1x10^6, 1x10^7, 1x10^8, 1x10^9, or 1x10^10 CiPSCs, intermediate plastic state cells, or somatic cells, or any combination thereof.

A composition may comprise a chemical reprogramming factor. A composition may comprise a plurality of chemical reprogramming factors. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more chemical reprogramming factors. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 chemical reprogramming factors. A composition may comprise 1 chemical reprogramming factors. A composition may comprise 2 chemical reprogramming factors. A composition may comprise 3 chemical reprogramming factors. A composition may comprise 4 chemical reprogramming factors. A composition may comprise 5 chemical reprogramming factors. A composition may comprise 6 chemical reprogramming factors. A composition may comprise 7 chemical reprogramming factors. A composition may comprise 8 chemical reprogramming factors. A composition may comprise 9 chemical reprogramming factors. A composition may comprise 10 chemical reprogramming factors. A composition may comprise 11 chemical reprogramming factors. A composition may comprise 12 chemical reprogramming factors. A composition may comprise 13 chemical reprogramming factors. A composition may comprise 14 chemical reprogramming factors. A composition may comprise 15 chemical reprogramming factors.

A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor, or any combination thereof. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, and a SAH hydrolase inhibitor. A composition may comprise a MEK inhibitor. A composition may comprise a B-Raf inhibitor. A composition may comprise a histone deacetylase inhibitor. A composition may comprise a Wnt inhibitor. A composition may comprise a GSK inhibitor. A composition may comprise a ROCK inhibitor. A composition may comprise an inhibitor of histone demethylation. A composition may comprise a Dot1L inhibitor. A composition may comprise a SAH hydrolase inhibitor.

A composition may comprise a MEK inhibitor, a B-Raf inhibitor, or a histone deacetylase inhibitor. A composition may comprise a MEK inhibitor or a B-Raf inhibitor. A composition may comprise a MEK inhibitor or a histone deacetylase inhibitor. A composition may comprise a B-Raf inhibitor or a histone deacetylase inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor.

A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a Wnt inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a GSK inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a ROCK inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and an inhibitor of histone demethylation. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a Dot1L inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a SAH hydrolase inhibitor. A composition may comprise a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor; and one or more of a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor.

A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor, or any combination thereof. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and at least 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and at most 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, and a SAH hydrolase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a B-Raf inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a histone deacetylase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a Wnt inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a GSK inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a ROCK inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and an inhibitor of histone demethylation. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a Dot1L inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a SAH hydrolase inhibitor. The intermediate plastic state cells may further express one or more of MSX2, NMYC, WNT4, FGF19, or TOP2A. Additionally, the intermediate plastic state cells may further express one or more of FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.

A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, or a histone deacetylase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor or a B-Raf inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor or a histone deacetylase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a B-Raf inhibitor or a histone deacetylase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor.

A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor, and a Wnt inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a GSK inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a ROCK inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and an inhibitor of histone demethylation. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a Dot1L inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a SAH hydrolase inhibitor. A composition may comprise intermediate plastic state cells that express LIN28A and SALL4; a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor; and one or more of a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor.

A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor, or any combination thereof. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and at least 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and at most 1, 2, 3, 4, 5, 6, 7, or 8 of a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, and a SAH hydrolase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a B-Raf inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a histone deacetylase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a Wnt inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a GSK inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a ROCK inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and an inhibitor of histone demethylation. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a Dot1L inhibitor.

A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, or a histone deacetylase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor or a B-Raf inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor or a histone deacetylase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a B-Raf inhibitor or a histone deacetylase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor.

A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a Wnt inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a GSK inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a ROCK inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and an inhibitor of histone demethylation. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor; and a Dot1L inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, and a SAH hydrolase inhibitor. A composition may comprise CiPSCs that express OCT4, SOX2, or NANOG; and a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor; and one or more of a Wnt inhibitor, a GSK inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor.

The CiPSCs of the compositions may further express one or more of FGF4, ZFP57, DPPA5, or REX1. Additionally, the CiPSCs of the compositions also express one or more of DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DNMT3L, or UTF1.

FIG. 6 depicts exemplary compositions comprising chemical reprogramming factors and optional cells. In FIG. 6, "A" represents the combination of a MEK inhibitor, a B-Raf inhibitor, and a histone deacetylase inhibitor; "B" represents an inhibitor of histone demethylation, a Dot1L inhibitor, or a SAH hydrolase inhibitor, or any combination thereof, the composition may or may not have any of the compounds in group B; "C" represents a Wnt inhibitor, a GSK inhibitor, or a ROCK inhibitor, any combination thereof, the composition may or may not have any of the compounds in group C; "D" represents intermediate plastic state cells, or CIPSCs, somatic cells, or any combination thereof, the composition may or may not have any of the cells in group D. For example, AB1C1D2 represents a composition that includes a MEK inhibitor, a B-Raf inhibitor, a histone deacetylase inhibitor, an inhibitor of histone demethylation, and CIPSCs.

A composition may comprise PD0325901, AZD8330, or TAK-733; SB590885, Vemurafenib, RAF265, and PLX4720; VPA, LMK235, MS275, or HDACi IV; IWR-1 or IWP-2; CHIR99021 or CHIR98014; Y-27632 or thiazovivin; Tranylcypromine; EPZ004777 or EPZ5676; DZNep, NepA, Adox, or DZA; or any combination thereof. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, or 8 of PD0325901, AZD8330, or TAK-733; SB590885, Vemurafenib, RAF265, and PLX4720; VPA, LMK235, MS275, or HDACi IV; IWR-1 or IWP-2; CHIR99021 or CHIR98014; Y-27632 or thiazovivin; Tranylcypromine; EPZ004777 or EPZ5676; or DZNep, NepA, Adox, or DZA. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, or 8 of PD0325901, AZD8330, or TAK-733; SB590885, Vemurafenib, RAF265, and PLX4720; VPA, LMK235, MS275, or HDACi IV; IWR-1 or IWP-2; CHIR99021 or CHIR98014; Y-27632 or thiazovivin; Tranylcypromine; EPZ004777 or EPZ5676; or DZNep, NepA, Adox, or DZA. A composition may comprise PD0325901, AZD8330, or TAK-733; SB590885, Vemurafenib, RAF265, and PLX4720; VPA, LMK235, MS275, or HDACi IV; IWR-1 or IWP-2; CHIR99021 or CHIR98014; Y-27632 or thiazovivin; Tranylcypromine; EPZ004777 or EPZ5676; and DZNep, NepA, Adox, or DZA. A composition may comprise PD0325901, AZD8330, or TAK-733. A composition may comprise SB590885, Vemurafenib, RAF265, and PLX4720. A composition may comprise VPA, LMK235, MS275, or HDACi IV. A composition may comprise IWR-1 or IWP-2. A composition may comprise CHIR99021 or CHIR98014. A composition may comprise Y-27632 or thiazovivin. A composition may comprise Tranylcypromine. A composition may comprise EPZ004777 or EPZ5676. A composition may comprise DZNep, NepA, Adox, or DZA.

A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, or VPA, LMK235, MS275, or HDACi IV. A composition may comprise PD0325901, AZD8330, or TAK-733 or SB590885, Vemurafenib, RAF265, and PLX4720. A composition may comprise PD0325901, AZD8330, or TAK-733 or VPA, LMK235, MS275, or HDACi IV. A composition may comprise SB590885, Vemurafenib, RAF265, and PLX4720 or VPA, LMK235, MS275, or HDACi IV. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV.

A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and IWR-1 or IWP-2. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and CHIR99021 or CHIR98014. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and Y-27632 or thiazovivin. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and Tranylcypromine. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and EPZ004777 or EPZ5676. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and DZNep, NepA, Adox, or DZA. A composition may comprise PD0325901, AZD8330, or TAK-733, SB590885, Vemurafenib, RAF265, and PLX4720, and VPA, LMK235, MS275, or HDACi IV; and one or more of IWR-1 or IWP-2, CHIR99021 or CHIR98014, Y-27632 or thiazovivin, Tranylcypromine, EPZ004777 or EPZ5676, or DZNep, NepA, Adox, or DZA.

A composition may comprise PD0325901; SB590885; VPA; IWP-2; CHIR99021; Y-27632; Tranylcypromine; EPZ004777; or DZNep; or any combination thereof. A composition may comprise at least 1, 2, 3, 4, 5, 6, 7, or 8 of PD0325901; SB590885; VPA; IWP-2; CHIR99021; Y-27632; Tranylcypromine; EPZ004777; or DZNep. A composition may comprise at most 1, 2, 3, 4, 5, 6, 7, or 8 of PD0325901; SB590885; VPA; IWP-2; CHIR99021; Y-27632; Tranylcypromine; EPZ004777; or DZNep. A composition may comprise PD0325901; SB590885; VPA; IWP-2; CHIR99021; Y-27632; Tranylcypromine; EPZ004777; and DZNep. A composition may comprise PD0325901. A composition may comprise SB590885. A composition may comprise VPA. A composition may comprise IWP-2. A composition may comprise CHIR99021. A composition may comprise Y-27632. A composition may comprise Tranylcypromine. A composition may comprise EPZ004777. A composition may comprise DZNep.

A composition may comprise PD0325901, SB590885, or VPA. A composition may comprise PD0325901 or SB590885. A composition may comprise PD0325901 or VPA. A composition may comprise SB590885 or VPA. A composition may comprise PD0325901, SB590885, and VPA.

A composition may comprise PD0325901, SB590885, and VPA; and IWP-2. A composition may comprise PD0325901, SB590885, and VPA; and CHIR99021. A composition may comprise PD0325901, SB590885, and VPA; and Y-27632. A composition may comprise PD0325901, SB590885, and VPA; and Tranylcypromine. A composition may comprise PD0325901, SB590885, and VPA; and EPZ004777. A composition may comprise PD0325901, SB590885, and VPA; and DZNep. A composition may comprise PD0325901, SB590885, and VPA; and one or more of IWP-2, CHIR99021, Y-27632, Tranylcypromine, EPZ004777, or DZNep.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 25 μM, 30 μM, 45 μM, 50 μM or more PD0325901 within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 25 μM, 30 μM, 45 μM, or 50 μM PD0325901 within the composition. A composition may comprise about 0.1 μM PD0325901 within the composition. A composition may comprise about 0.2 μM PD0325901 within the composition. A composition may comprise about 0.3 μM PD0325901 within the composition. A composition may comprise about 0.4 μM PD0325901 within the composition. A composition may comprise about 0.5 μM PD0325901 within the composition. A composition may comprise about 0.6 μM PD0325901 within the composition. A composition may comprise about 0.7 μM PD0325901 within the composition. A composition may comprise about 0.8 μM PD0325901 within the composition. A composition may comprise about 0.9 μM PD0325901 within the composition. A composition may comprise about 1 μM PD0325901 within the composition. A composition may comprise about 2 μM PD0325901 within the composition. A composition may comprise about 3 μM PD0325901 within the composition. A composition may comprise about 4 μM PD0325901 within the composition. A composition may comprise about 5 μM PD0325901 within the composition. A composition may comprise about 6 μM PD0325901 within the composition. A composition may comprise about 7 μM PD0325901 within the composition. A composition may comprise about 8 μM PD0325901 within the composition. A composition may comprise about 9 μM PD0325901 within the composition. A composition may comprise about 10 μM PD0325901 within the composition. A composition may comprise about 0.1 μM to about 10 μM PD0325901 within the composition. A composition may comprise about 0.2 μM to about 7.5 μM PD0325901 within the composition. A composition may comprise about 0.3 μM to about 5 μM PD0325901 within the composition. A composition may comprise about 0.4 μM to about 4 μM PD0325901 within the composition. A composition may comprise about 0.5 μM to about 3 μM PD0325901 within the composition. A composition may comprise about 0.75 μM to about 2 μM PD0325901 within the composition.

A composition may comprise at least about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, 25 μM or more SB590885 within the composition. A composition may comprise at most about 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.75 μM, 1 μM, 1.25 μM, 1.5 μM, 1.75 μM, 2 μM, 2.25 μM, 2.5 μM, 2.75 μM, 3 μM, 3.25 μM, 3.5 μM, 3.75 μM, 4 μM, 4.25 μM, 4.5 μM, 4.75 μM, 5 μM, 7.5 μM, 10 μM, 12.5 μM, 15 μM, 17.5 μM, 20 μM, 22.5 μM, or 25 μM SB590885 within the composition. A composition may comprise about 0.05 μM SB590885 within the composition. A composition may comprise about 0.1 μM SB590885 within the composition. A composition may comprise about 0.15 μM SB590885 within the composition. A composition may comprise about 0.2 μM SB590885 within the composition. A composition may comprise about 0.25 μM SB590885 within the composition. A composition may comprise about 0.3 μM SB590885 within the composition. A composition may comprise about 0.35 μM SB590885 within the composition. A composition may comprise about 0.4 μM SB590885 within the composition. A composition may comprise about 0.45 μM SB590885 within the composition. A composition may comprise about 0.5 μM SB590885 within the composition. A composition may comprise about 1 μM SB590885 within the composition. A composition may comprise about 1.5 μM SB590885 within the composition. A composition may comprise about 2 μM SB590885 within the composition. A composition may comprise about 2.5 μM SB590885 within the composition. A composition may comprise about 3 μM SB590885 within the composition. A composition may comprise about 3.5 μM SB590885 within the composition. A composition may comprise about 4 μM SB590885 within the composition. A composition may comprise about 4.5 μM SB590885 within the composition. A composition may comprise about 5 μM SB590885 within the composition. A composition may comprise about 0.05 μM to about 2.5 μM SB590885 within the composition. A composition may comprise about 0.1 μM to about 1.875 μM SB590885 within the composition. A composition may comprise about 0.15 μM to about 1.25 μM SB590885 within the composition. A composition may comprise about 0.2 μM to about 1 μM SB590885 within the composition. A composition may comprise about 0.25 μM to about 0.75 μM SB590885 within the composition. A composition may comprise about 0.375 μM to about 0.5 μM SB590885 within the composition.

A composition may comprise at least about 0.02 millimolar (mM) , 0.04 mM, 0.06 mM, 0.08 mM, 0.1 mM, 0.12 mM, 0.14 mM, 0.16 mM, 0.18 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 25 mM, 30 mM, 45 mM, 50 mM or more VPA within the composition. A composition may comprise at most about 0.02 mM, 0.04 mM, 0.06 mM, 0.08 mM, 0.1 mM, 0.12 mM, 0.14 mM, 0.16 mM, 0.18 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 25 mM, 30 mM, 45 mM, or 50 mM VPA within the composition. A composition may comprise about 0.1 mM VPA within the composition. A composition may comprise about 0.2 mM VPA within the composition. A composition may comprise about 0.3 mM VPA within the composition. A composition may comprise about 0.4 mM VPA within the composition. A composition may comprise about 0.5 mM VPA within the composition. A composition may comprise about 0.6 mM VPA within the composition. A composition may comprise about 0.7 mM VPA within the composition. A composition may comprise about 0.8 mM VPA within the composition. A composition may comprise about 0.9 mM VPA within the composition. A composition may comprise about 1 mM VPA within the composition. A composition may comprise about 2 mM VPA within the composition. A composition may comprise about 3 mM VPA within the composition. A composition may comprise about 4 mM VPA within the composition. A composition may comprise about 5 mM VPA within the composition. A composition may comprise about 6 mM VPA within the composition. A composition may comprise about 7 mM VPA within the composition. A composition may comprise about 8 mM VPA within the composition. A composition may comprise about 9 mM VPA within the composition. A composition may comprise about 10 mM VPA within the composition. A composition may comprise about 0.1 mM to about 10 mM VPA within the composition. A composition may comprise about 0.2 mM to about 7.5 mM VPA within the composition. A composition may comprise about 0.3 mM to about 5 mM VPA within the composition. A composition may comprise about 0.4 mM to about 4 mM VPA within the composition. A composition may comprise about 0.5 mM to about 3 mM VPA within the composition. A composition may comprise about 0.75 mM to about 2 mM VPA within the composition.

A composition may comprise at least about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, 500 μM or more Tranylcypromine within the composition. A composition may comprise at most about 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 21 μM, 22 μM, 23 μM, 24 μM, 25 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, 200 μM, 300 μM, 400 μM, or 500 μM Tranylcypromine within the composition. A composition may comprise about 1 μM Tranylcypromine within the composition. A composition may comprise about 2 μM Tranylcypromine within the composition. A composition may comprise about 3 μM Tranylcypromine within the composition. A composition may comprise about 4 μM Tranylcypromine within the composition. A composition may comprise about 5 μM Tranylcypromine within the composition. A composition may comprise about 6 μM Tranylcypromine within the composition. A composition may comprise about 7 μM Tranylcypromine within the composition. A composition may comprise about 8 μM Tranylcypromine within the composition. A composition may comprise about 9 μM Tranylcypromine within the composition. A composition may comprise about 10 μM Tranylcypromine within the composition. A composition may comprise about 15 μM Tranylcypromine within the composition. A composition may comprise about 20 μM Tranylcypromine within the composition. A composition may comprise about 30 μM Tranylcypromine within the composition. A composition may comprise about 40 μM Tranylcypromine within the composition. A composition may comprise about 50 μM Tranylcypromine within the composition. A composition may comprise about 60 μM Tranylcypromine within the composition. A composition may comprise about 70 μM Tranylcypromine within the composition. A composition may comprise about 80 μM Tranylcypromine within the composition. A composition may comprise about 90 μM Tranylcypromine within the composition. A composition may comprise about 100 μM Tranylcypromine within the composition. A composition may comprise about 1 μM to about 100 μM Tranylcypromine within the composition. A composition may comprise about 2 μM to about 75 μM Tranylcypromine within the composition. A composition may comprise about 3 μM to about 50 μM Tranylcypromine within the composition. A composition may comprise about 4 μM to about 40 μM Tranylcypromine within the composition. A composition may comprise about 5 μM to about 30 μM Tranylcypromine within the composition. A composition may comprise about 7.5 μM to about 20 μM Tranylcypromine within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more EPZ5676 within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM EPZ5676 within the composition. A composition may comprise about 0.2 μM EPZ5676 within the composition. A composition may comprise about 0.4 μM EPZ5676 within the composition. A composition may comprise about 0.6 μM EPZ5676 within the composition. A composition may comprise about 0.8 μM EPZ5676 within the composition. A composition may comprise about 1 μM EPZ5676 within the composition. A composition may comprise about 1.2 μM EPZ5676 within the composition. A composition may comprise about 1.4 μM EPZ5676 within the composition. A composition may comprise about 1.6 μM EPZ5676 within the composition. A composition may comprise about 1.8 μM EPZ5676 within the composition. A composition may comprise about 2 μM EPZ5676 within the composition. A composition may comprise about 4 μM EPZ5676 within the composition. A composition may comprise about 6 μM EPZ5676 within the composition. A composition may comprise about 8 μM EPZ5676 within the composition. A composition may comprise about 10 μM EPZ5676 within the composition. A composition may comprise about 12 μM EPZ5676 within the composition. A composition may comprise about 14 μM EPZ5676 within the composition. A composition may comprise about 16 μM EPZ5676 within the composition. A composition may comprise about 18 μM EPZ5676 within the composition. A composition may comprise about 20 μM EPZ5676 within the composition. A composition may comprise about 0.2 μM to about 20 μM EPZ5676 within the composition. A composition may comprise about 0.4 μM to about 15 μM EPZ5676 within the composition. A composition may comprise about 0.6 μM to about 10 μM EPZ5676 within the composition. A composition may comprise about 0.8 μM to about 8 μM EPZ5676 within the composition. A composition may comprise about 1 μM to about 6 μM EPZ5676 within the composition. A composition may comprise about 1.5 μM to about 4 μM EPZ5676 within the composition.

A composition may comprise at least about 0.0004 μM, 0.0008 μM, 0.0012 μM, 0.0016 μM, 0.002 μM, 0.004 μM, 0.006 μM, 0.008 μM, 0.01 μM, 0.012 μM, 0.014 μM, 0.016 μM, 0.018 μM, 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1 μM or more DZNep within the composition. A composition may comprise at most about 0.0004 μM, 0.0008 μM, 0.0012 μM, 0.0016 μM, 0.002 μM, 0.004 μM, 0.006 μM, 0.008 μM, 0.01 μM, 0.012 μM, 0.014 μM, 0.016 μM, 0.018 μM, 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.14 μM, 0.16 μM, 0.18 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, or 1 μM DZNep within the composition. A composition may comprise about 0.002 μM DZNep within the composition. A composition may comprise about 0.004 μM DZNep within the composition. A composition may comprise about 0.006 μM DZNep within the composition. A composition may comprise about 0.008 μM DZNep within the composition. A composition may comprise about 0.01 μM DZNep within the composition. A composition may comprise about 0.012 μM DZNep within the composition. A composition may comprise about 0.014 μM DZNep within the composition. A composition may comprise about 0.016 μM DZNep within the composition. A composition may comprise about 0.018 μM DZNep within the composition. A composition may comprise about 0.02 μM DZNep within the composition. A composition may comprise about 0.04 μM DZNep within the composition. A composition may comprise about 0.06 μM DZNep within the composition. A composition may comprise about 0.08 μM DZNep within the composition. A composition may comprise about 0.1 μM DZNep within the composition. A composition may comprise about 0.12 μM DZNep within the composition. A composition may comprise about 0.14 μM DZNep within the composition. A composition may comprise about 0.16 μM DZNep within the composition. A composition may comprise about 0.18 μM DZNep within the composition. A composition may comprise about 0.2 μM DZNep within the composition. A composition may comprise about 0.002 μM to about 0.2 μM DZNep within the composition. A composition may comprise about 0.0025 μM to about 0.15 μM DZNep within the composition. A composition may comprise about 0.005 μM to about 0.1 μM DZNep within the composition. A composition may comprise about 0.0075 μM to about 0.75 μM DZNep within the composition. A composition may comprise about 0.01 μM to about 0.5 μM DZNep within the composition. A composition may comprise about 0.015 μM to about 0.4 μM DZNep within the composition.

A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM or more IWP-2 within the composition. A composition may comprise at least about 0.04 μM, 0.08 μM, 0.12 μM, 0.16 μM, 0.2 μM, 0.4 μM, 0.6 μM, 0.8 μM, 1 μM, 1.2 μM, 1.4 μM, 1.6 μM, 1.8 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, 10 μM, 11 μM, 12 μM, 13 μM, 14 μM, 15 μM, 16 μM, 17 μM, 18 μM, 19 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, or 100 μM IWP-2 within the composition. A composition may comprise about 0.2 μM IWP-2 within the composition. A composition may comprise about 0.4 μM IWP-2 within the composition. A composition may comprise about 0.6 μM IWP-2 within the composition. A composition may comprise about 0.8 μM IWP-2 within the composition. A composition may comprise about 1 μM IWP-2 within the composition. A composition may comprise about 1.2 μM IWP-2 within the composition. A composition may comprise about 1.4 μM IWP-2 within the composition. A composition may comprise about 1.6 μM IWP-2 within the composition. A composition may comprise about 1.8 μM IWP-2 within the composition. A composition may comprise about 2 μM IWP-2 within the composition. A composition may comprise about 4 μM IWP-2 within the composition. A composition may comprise about 6 μM IWP-2 within the composition. A composition may comprise about 8 μM IWP-2 within the composition. A composition may comprise about 10 μM IWP-2 within the composition. A composition may comprise about 12 μM IWP-2 within the composition. A composition may comprise about 14 μM IWP-2 within the composition. A composition may comprise about 16 μM IWP-2 within the composition. A composition may comprise about 18 μM IWP-2 within the composition. A composition may comprise about 20 μM IWP-2 within the composition. A composition may comprise about 0.2 μM to about 20 μM IWP-2 within the composition. A composition may comprise about 0.4 μM to about 15 μM IWP-2 within the composition. A composition may comprise about 0.6 μM to about 10 μM IWP-2 within the composition. A composition may comprise about 0.8 μM to about 8 μM IWP-2 within the composition. A composition may comprise about 1 μM to about 6 μM IWP-2 within the composition. A composition may comprise about 1.5 μM to about 4 μM IWP-2 within the composition.

A composition may comprise at least about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, 50 μM or more CHIR99021 within the composition. A composition may comprise at most about 0.02 μM, 0.04 μM, 0.06 μM, 0.08 μM, 0.1 μM, 0.2 μM, 0.3 μM, 0.4 μM, 0.5 μM, 0.6 μM, 0.7 μM, 0.8 μM, 0.9 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, 40 μM, 45 μM, or 50 μM CHIR99021 within the composition. A composition may comprise about 0.1 μM CHIR99021 within the composition. A composition may comprise about 0.2 μM CHIR99021 within the composition. A composition may comprise about 0.3 μM CHIR99021 within the composition. A composition may comprise about 0.4 μM CHIR99021 within the composition. A composition may comprise about 0.5 μM CHIR99021 within the composition. A composition may comprise about 0.6 μM CHIR99021 within the composition. A composition may comprise about 0.7 μM CHIR99021 within the composition. A composition may comprise about 0.8 μM CHIR99021 within the composition. A composition may comprise about 0.9 μM CHIR99021 within the composition. A composition may comprise about 1 μM CHIR99021 within the composition. A composition may comprise about 2 μM CHIR99021 within the composition. A composition may comprise about 3 μM CHIR99021 within the composition. A composition may comprise about 4 μM CHIR99021 within the composition. A composition may comprise about 5 μM CHIR99021 within the composition. A composition may comprise about 6 μM CHIR99021 within the composition. A composition may comprise about 7 μM CHIR99021 within the composition. A composition may comprise about 8 μM CHIR99021 within the composition. A composition may comprise about 9 μM CHIR99021 within the composition. A composition may comprise about 10 μM CHIR99021 within the composition. A composition may comprise about 0.1 μM to about 10 μM CHIR99021 within the composition. A composition may comprise about 0.2 μM to about 7.5 μM CHIR99021 within the composition. A composition may comprise about 0.3 μM to about 5 μM CHIR99021 within the composition. A composition may comprise about 0.4 μM to about 4 μM CHIR99021 within the composition. A composition may comprise about 0.5 μM to about 3 μM CHIR99021 within the composition. A composition may comprise about 0.75 μM to about 2 μM CHIR99021 within the composition.

Subsequent to contacting any populations of stage 2 cells with any compositions described herein, the cells may be incubated in normoxic condition. For example, the stage 2 cells may be incubated with at most 23%, 22%, 21%, 20%, or 19%atmospheric oxygen. The normoxic condition may comprise about 22%atmospheric oxygen. The normoxic condition may comprise about 21%atmospheric oxygen. The normoxic condition may comprise about 20%atmospheric oxygen.

Subsequent to contacting any populations of stage 2 cells with any compositions described herein, a population of stage 2 cells may be incubated with a composition for at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 25 days, or 30 days. A population of stage 2 cells may be incubated with a composition for at most about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 25 days, or 30 days. A population of stage 2 cells may be incubated with a composition for about 1 day. A population of stage 2 cells may be incubated with a composition for about 2 days. A population of stage 2 cells may be incubated with a composition for about 3 days. A population of stage 2 cells may be incubated with a composition for about 4 days. A population of stage 2 cells may be incubated with a composition for about 5 days. A population of stage 2 cells may be incubated with a composition for about 6 days. A population of stage 2 cells may be incubated with a composition for about 7 days. A population of stage 2 cells may be incubated with a composition for about 8 days. A population of stage 2 cells may be incubated with a composition for about 9 days. A population of stage 2 cells may be incubated with a composition for about 10 days. A population of stage 2 cells may be incubated with a composition for about 11 days. A population of stage 2 cells may be incubated with a composition for about 12 days. A population of stage 2 cells may be incubated with a composition for about 13 days. A population of stage 2 cells may be incubated with a composition for about 14 days. A population of stage 2 cells may be incubated with a composition for about 15 days. A population of stage 2 cells may be incubated with a composition for about 16 days. A population of stage 2 cells may be incubated with a composition for about 17 days. A population of stage 2 cells may be incubated with a composition for about 18 days. A population of stage 2 cells may be incubated with a composition for about 19 days. A population of stage 2 cells may be incubated with a composition for about 20 days. A population of stage 2 cells may be incubated with a composition for about 25 days.

In some cases, subsequent to incubating a population of stage 2 cells with a first composition for a first period of time, the first composition may be removed from the cells. The cells can then be contacted with a second composition. Subsequent to incubating the cells with the second composition for a second period of time, the second composition may be removed from the cells. The cells can then be contacted with a third composition and incubated with the third composition for a third period of time. The first period of time may comprise at least about 1, 2, 3, 4, 5, 6, 7, or 8 days. The first period of time may comprise at most about 1, 2, 3, 4, 5, 6, 7, or 8 days. The second period of time may comprise at least about 1, 2, 3, 4, 5, 6, 7, or 8 days. The second period of time may comprise at most about 1, 2, 3, 4, 5, 6, 7, or 8 days. The third period of time may comprise at least about 1, 2, 3, 4, 5, 6, 7, or 8 days. The third period of time may comprise at most about 1, 2, 3, 4, 5, 6, 7, or 8 days. The first composition may comprise the histone deacetylase inhibitor, the inhibitor of histone demethylation, the SAH hydrolase inhibitor, or the Dot1L inhibitor. The second composition may comprise the histone deacetylase inhibitor with the amount that is at least about 30%, 40%, 50%, 60%, or 70%less than the first composition; the inhibitor of histone demethylation; the SAH hydrolase inhibitor; or the Dot1L inhibitor. The second composition may comprise the histone deacetylase inhibitor with the amount that is at most about 30%, 40%, 50%, 60%, or 70%less than the first composition; the inhibitor of histone demethylation; the SAH hydrolase inhibitor; or the Dot1L inhibitor. The third composition may not comprise the inhibitor of histone demethylation; the SAH hydrolase inhibitor; or the Dot1L inhibitor.

CHEMICAL REPROGRAMMING FACTORS

Compositions provided herein may comprise at least a cell or at least a chemical reprogramming factor. In some cases, a composition provided herein comprises a cell or a population of cells. In some cases, a composition provided herein comprises a chemical reprogramming factor. A composition may comprise at least a chemical reprogramming factor. A composition may comprise a plurality of chemical reprogramming factors. A composition may comprise at least a cell and at least a chemical reprogramming factor. A composition may comprise a population of cells and a plurality of chemical reprogramming factors.

In some cases, the compositions described herein comprise a chemical reprogramming factor. A chemical reprogramming factor may facilitate a conversion of a cell type to another cell type. A chemical reprogramming factor may facilitate a cell conversion by increasing the number of reprogrammed cells obtained from the same starting cell density cultured for the same length of time and/or improving the quality of reprogrammed cells, measured in terms of characteristics selected from the ability of the cells to express pluripotency factors such as OCT4, SOX2 and NANOG and number of passages in culture, when compared to a reprograming method that does not use the same chemical reprogramming factor.

A chemical reprogramming factor may regulate a cellular component involved in a cellular activity or biological activity. In some cases, the compositions described herein comprise a plurality of chemical reprogramming factors. A chemical reprogramming factor may comprise a chemical compound that can facilitate the conversion of a first population of cells to a second population of cells. A chemical reprogramming factor may comprise a chemical compound that can facilitate the conversion of a cell type to another cell type. A chemical reprogramming factor may not induce a genetic modification a cell. A genetic modification may comprise a change in the make-up of a genome of a cell. The genome may comprise a chromosomal deoxyribonucleic acid (DNA) or an extra-chromosomal DNA (e.g., a mitochondrial DNA) . A genetic modification may comprise a mutation. The mutation may comprise a substitution, deletion, or insertion of the genome. In other cases, the mutation also comprises a DNA rearrangement of the chromosomal or mitochondrial DNA. A chemical reprogramming factor may not add exogenous genetic materials into a cell. A chemical reprogramming factor may not add exogenous genetic materials a chromosomal or extra- chromosomal DNA of the cell.

A chemical reprogramming factor may comprise a chemical compound or molecule. A chemical reprogramming factor may comprise an organic compound or an inorganic compound. A chemical reprogramming factor may comprise an organic compound. A chemical reprogramming factor may comprise an inorganic compound. A chemical reprogramming factor may be peptide-based. A chemical reprogramming factor may not be peptide-based. A chemical reprogramming factor may comprise a carbohydrate moiety. A chemical reprogramming factor may not comprise a carbohydrate moiety. A chemical reprogramming factor may comprise a lipid moiety. A chemical reprogramming factor may not comprise a lipid moiety. A chemical reprogramming factor may comprise a drug. In some cases, A chemical reprogramming factor may comprise any chemical reprogramming factors described herein. In some cases, a chemical reprogramming factor is substituted with another chemical reprogramming factor described herein. In some cases, a chemical reprogramming factor is substituted with another chemical reprogramming factor identified based on the methods described herein.

A chemical reprogramming factor may elicit a biological activity of a cell when the cell contacted the chemical reprogramming factor. A biological activity may comprise a biophysical or biochemical response of a cell. Such a biophysical response may comprise cell locomotion, cell attachment/detachment, cell polarization, cell shape change, or any combination thereof. A biochemical response may comprise gene expression, protein expression, RNA expression, post-transcriptional modification of a protein/DNA/ribonucleic acid (RNA) , post-translational modification of a protein, endocytosis, exocytosis, cell proliferation, cell-cycle progression, cell differentiation, or any combination thereof. In some cases, a biological activity comprises a cellular activity described herein.

In some cases, two chemical reprogramming factors share a same biological activity. To measure a biological activity of a chemical reprogramming factor, a biological assay may be used. A biological assay may qualitatively or quantitatively reflect or report a biological activity to be measured. The specific assay used for a specific biological activity may depend on the biological activity being measure. The biological assay may be an in vitro assay. The biological assay may be an in vivo assay. The specific assay may derive a measurable value of the biological activity of a chemical reprogramming factor. In some cases, two chemical reprogramming factors are determined to share a same biological activity if the difference of their measurable values in a same biological assay is within 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, 4-fold, or 5-fold. In some cases, two chemical reprogramming factors determined to share a same biological activity may substitute each other for converting a cell. The measurable value may be derived from various markers involved in various biological activity described herein.

A chemical reprogramming factor may have a molecular weight. The molecular weight of a chemical reprogramming factor may be at most about 10000 Daltons (Da) , 9000 Da, 8000 Da, 7000 Da, 6000 Da, 5000 Da, 4500 Da, 4000 Da, 3500 Da, 3000 Da, 2500 Da, 2000 Da, 1900 Da, 1800 Da, 1700 Da, 1600 Da, 1500 Da, 1400 Da, 1300 Da, 1200 Da, 1100 Da, 1000 Da, 900 Da, 800 Da, 700 Da, 600 Da, 500 Da, 400 Da, 300 Da, 200 Da, 100 Da or less. The molecular weight of a chemical reprogramming factor may be at less about 100 Da, 200 Da, 300 Da, 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1000 Da, 1100 Da, 1200 Da, 1300 Da, 1400 Da, 1500 Da, 1600 Da, 1700 Da, 1800 Da, 1900 Da, 2000 Da, 2500 Da, 3000 Da, 3500 Da, 4000 Da, 4500 Da, 5000 Da, 6000 Da, 7000 Da, 8000 Da, 9000 Da, 10000 Da or more. The molecular weight of a chemical reprogramming factor may be at least about 100 Da. The molecular weight of a chemical reprogramming factor may be at least about 200 Da. The molecular weight of a chemical reprogramming factor may be at least about 300 Da. The molecular weight of a chemical reprogramming factor may be at least about 400 Da. The molecular weight of a chemical reprogramming factor may be at least about 500 Da. The molecular weight of a chemical reprogramming factor may be at least about 600 Da. The molecular weight of a chemical reprogramming factor may be at least about 700 Da. The molecular weight of a chemical reprogramming factor may be at least about 800 Da. The molecular weight of a chemical reprogramming factor may be at least about 900 Da. The molecular weight of a chemical reprogramming factor may be at least about 1000 Da. The molecular weight of a chemical reprogramming factor may be at least about 1100 Da. The molecular weight of a chemical reprogramming factor may be at least about 1200 Da. The molecular weight of a chemical reprogramming factor may be at least about 1300 Da. The molecular weight of a chemical reprogramming factor may be at least about 1400 Da. The molecular weight of a chemical reprogramming factor may be at least about 1500 Da. The molecular weight of a chemical reprogramming factor may be at least about 1600 Da. The molecular weight of a chemical reprogramming factor may be at least about 1700 Da. The molecular weight of a chemical reprogramming factor may be at least about 1800 Da. The molecular weight of a chemical reprogramming factor may be at least about 1900 Da. The molecular weight of a chemical reprogramming factor may be at least about 2000 Da. The molecular weight of a chemical reprogramming factor may be at most about 100 Da. The molecular weight of a chemical reprogramming factor may be at most about 200 Da. The molecular weight of a chemical reprogramming factor may be at most about 300 Da. The molecular weight of a chemical reprogramming factor may be at most about 400 Da. The molecular weight of a chemical reprogramming factor may be at most about 500 Da. The molecular weight of a chemical reprogramming factor may be at most about 600 Da. The molecular weight of a chemical reprogramming factor may be at most about 700 Da. The molecular weight of a chemical reprogramming factor may be at most about 800 Da. The molecular weight of a chemical reprogramming factor may be at most about 900 Da. The molecular weight of a chemical reprogramming factor may be at most about 1000 Da. The molecular weight of a chemical reprogramming factor may be at most about 1100 Da. The molecular weight of a chemical reprogramming factor may be at most about 1200 Da. The molecular weight of a chemical reprogramming factor may be at most about 1300 Da. The molecular weight of a chemical reprogramming factor may be at most about 1400 Da. The molecular weight of a chemical reprogramming factor may be at most about 1500 Da. The molecular weight of a chemical reprogramming factor may be at most about 1600 Da. The molecular weight of a chemical reprogramming factor may be at most about 1700 Da. The molecular weight of a chemical reprogramming factor may be at most about 1800 Da. The molecular weight of a chemical reprogramming factor may be at most about 1900 Da. The molecular weight of a chemical reprogramming factor may be at most about 2000 Da.

In some cases, a plurality of chemical reprogramming factors facilitates a conversion of a first population of cells to a second population of cells. In some cases, while a plurality of chemical reprogramming factors facilitates a conversion of a first population of cells to a second population of cells, a composition without any one of the chemical reprogramming factors may not facilitate the conversion of the first population of cells to the second populations of cells. In some cases, each of the plurality of chemical reprogramming factors has a dosage range to facilitate the conversion of the first population of cells to the second populations of cells. In some cases, if any one of the chemical reprogramming factors does not have the dosage range described herein, the plurality of chemical reprogramming factors may not facilitate the conversion of the first population of cells to the second populations of cells. In some cases, the dosage range for a chemical reprogramming factor for converting a population of cells may comprises any dosage ranges described herein. In some cases, the dosage range for a chemical reprogramming factor for converting a population of cells may comprises the dosage ranges identified using the methods described herein (e.g., using the methods described in the

EXAMPLEs described herein.

In some cases, two chemical reprogramming factors may substitute each other for converting a cell by testing the conversion or reprogramming efficiencies using methods described in the EXAMPLEs described herein.

Provided herein, in some aspects, are exemplary chemical reprogramming factors that may be used in a method provided herein or may be contained in a composition provided herein. These chemical reprogramming factors may increase or decrease various cellular or biological activities.

1. Glycogen synthesis kinase (GSK; or glycogen kinase) Inhibitors

A chemical reprogramming factor may comprise a chemical compound that can inhibit GSK. GSK may comprise a serine/threonine protein kinase that mediates phosphorylation of serine and threonine of various cellular factors. The phosphorylation of these cellular factors may control glycogen metabolism, cell signaling, or cellular transport. GSK inhibition may lead to a decrease in glycogen synthesis in the liver and muscles and/or increased blood glucose or hyperglycemia. GSK inhibition in neuroblastoma may reduce neuroendocrine marker expression (complex-like1 (ASCL1) and chromogranin A (CgA) , and/or beta-catenin) and/or suppress neuroblastoma cell growth (Carter et al., 2014; Cancer Biol Ther . 2014 May; 15 (5) : 510-5, which is herein incorporated by reference in its entirety) . GSK inhibition in cancer cells may decrease the growth of the cancer cell (Carter 2014) .

A GSK inhibitor may comprise CHIR99021 ( [6- [ [2- [ [4- (2, 4-Dichlorophenyl) -5- (5-methyl-1H-imidazol-2-yl) -2-pyrimidinyl] amino] ethyl] amino] -3-pyridinecarbonitrile] ) ; BIO-acetoxime; GSK 3I inhibitor XV; SB-216763 ( [3- (2, 4-Dichlorophenyl) -4- (1-methyl-1H-indol-3-yl) -1H-pyrrole-2, 5-dione] ) ; CHIR99021 trihydrochloride (ahydrochloride salt of CHIR99021) ; GSK-3 Inhibitor IX ( [ ( (2Z, 3E) -6’ -bromo-3- (hydroxyimino) - [2, 3’ -biindolinylidene] -2’ -one] ) ; GSK3 IX ( [6-Bromoindirubin-3'-oxime] ) ; GSK-3β Inhibitor XII ( [3- [ [6- (3-Aminophenyl) -7H-pyrrolo [2, 3-d] pyrimidin-4-yl] oxy] phenol] ) ; GSK-3 Inhibitor XVI ( [6- (2- (4- (2, 4-dichlorophenyl) -5- (4-methyl-1H-imidazol-2-yl) -pyrimidin-2-ylamino) ethyl-amino) -nicotinonitrile] ) ; SB-415286 ( [3- [ (3-chloro-4-hydroxyphenyl) amino] -4- (2-nitrophenyl) -1 H-pyrrole-2, 5-dione] ) ; Bio ( [ (2'Z, 3'E) -6-bromoindirubin-3'-oxime] ) ; TD114-2 ( [6, 7, 9, 10, 12, 13, 15, 16, 18, 19-Decahydro-5, 29: 20, 25-dimetheno-26H-dibenzo [n, t] pyrrolo [3, 4-q] [1, 4, 7, 10, 13, 22] tetraoxadiazacyclote tracosine-26, 28 (27H) -dione] ) ; or CHIR98014 ( [N6- [2- [ [4- (2, 4-Dichlorophenyl) -5- (1H-imidazol-1-yl) -2-pyrimidinyl] amino] ethyl] -3-nitro-2, 6-pyridinediamine] ) or any combination thereof. A GSK inhibitor may comprise A GSK inhibitor may comprise CHIR99021. A GSK inhibitor may comprise BIO-acetoxime. A GSK inhibitor may comprise GSK 3I inhibitor XV. A GSK inhibitor may comprise SB-216763. A GSK inhibitor may comprise CHIR99021 trihydrochloride. A GSK inhibitor may comprise GSK-3 Inhibitor IX. A GSK inhibitor may comprise GSK3 IX. A GSK inhibitor may comprise GSK-3β Inhibitor XII. A GSK inhibitor may comprise GSK-3 Inhibitor XVI. A GSK inhibitor may comprise SB-415286. A GSK inhibitor may comprise Bio. A GSK inhibitor may comprise TD114-2. A GSK inhibitor may comprise CHIR98014. aminopyrimidine. Aminopyrimidine may comprise CHIR99021, CHIR99021 trihydrochloride, or CHIR98014, or any combination thereof.

2. Transforming growth factor beta receptor inhibitors (TGFβR inhibitors)

A chemical reprogramming factor may comprise a chemical compound that can inhibit TGFβ receptor. The type I TGFβ receptor may comprise activin receptor-like kinase (ALK) 1, ALK2, ALK3, ALK4, ALK5, ALK6, or ALK7. The type II TGFβ receptor may comprise TGFβR2, bone morphogenetic protein receptor type 2 (BMPR2) , activin receptor type-2A (ACVR2A) , ACVR2B, or anti-Müllerian hormone receptor 2 (AMHR2) . The type III TGFβreceptor may comprise TGFβR3 (β-glycan) . A TGFβ receptor inhibitor may inhibit type I TGFβreceptor. A TGFβ receptor inhibitor may inhibit type II TGFβ receptor. A TGFβ receptor inhibitor may inhibit type III TGFβ receptor. A TGFβ receptor inhibitor may inhibit ALK1, ALK2, ALK3, ALK4, ALK5, ALK6, ALK7, BMPR2, ACVR2A, ACVR2B, AMHR2, TGFβR3, or any combination thereof. A TGFβ receptor inhibitor used in the subject methods or compositions may be an ALK inhibitor or a type I TGFβ receptor inhibitor, e.g., specifically inhibiting one or more type I TGFβ receptors, such as ALK1, ALK2, ALK3, ALK4, ALK5, ALK6, or ALK7. In some cases, a TGFβ receptor inhibitor used in the subject methods or compositions does not specifically inhibit TGFβ receptors other than type I TGFβ receptors, for instance, a TGFβ receptor inhibitor used in the subject methods or compositions may not inhibit any BMP receptor. In some cases, a TGFβ receptor inhibitor is an ALK5 inhibitor. In some cases, a TGFβ receptor inhibitor is an ALK5 inhibitor that does not specifically inhibit ALK2, ALK3, or ALK6.

A TGFβ receptor may be activated by growth factors or cytokines. Inhibition of the TGFβ receptor may lead to decreased cell proliferation or decreased metastasis of cancers (Derynck et al, 2003; Nature . 2003 Oct 9; 425 (6958) : 577-84., which is herein incorporated by reference in its entirety) . Inhibition of TGFβ receptor may lead to a decreased phosphorylation of R-Smad (or Receptor-regulated Mothers against decapentaplegic; Derynck 2003) . In some cases, Inhibition of TGFβ receptor may lead to a decreased transcription of genes under the regulation of Smad binding element (Derynck 2003) .

A TGFβ receptor inhibitor may comprise E-616452 (or 616452; [2- (3- (6-Methylpyridin-2-yl) -1H-pyrazol-4-yl) -1, 5-naphthyridine] ) ; A 83-01 ( [3- (6-Methyl-2-pyridinyl) -N-phenyl-4- (4-quinolinyl) -1H-pyrazole-1-carbothioamide] ; SB 505124 ( [2- [4- (1, 3-Benzodioxol-5-yl) -2- (1, 1-dimethylethyl) -1H-imidazol-5-yl] -6-methyl-pyridine] ) ; GW 788388 ( [4- [4- [3- (2-Pyridinyl) -1H-pyrazol-4-yl] -2-pyridinyl] -N- (tetrahydro-2H-pyran-4-yl) -benzamide] ) ; SB 525334 ( [6- [2- (1, 1-Dimethylethyl) -5- (6-methyl-2-pyridinyl) -1H-imidazol-4-yl] quinoxaline] ) , Dorsomorphin, or SB431542, or any combination thereof. A TGFβ receptor inhibitor may comprise E-616452. A TGFβ receptor inhibitor may comprise A 83-01. A TGFβreceptor inhibitor may comprise SB 505124. A TGFβ receptor inhibitor may comprise GW 788388. A TGFβ receptor inhibitor may comprise SB 525334. A TGFβ receptor inhibitor may comprise Dorsomorphin. A TGFβ receptor inhibitor may not comprise Dorsomorphin. A TGFβreceptor inhibitor may comprise SB431542.

In some cases, a chemical reprogramming factor may comprise a BMP receptor/AMPK inhibitor. A BMP receptor/AMPK inhibitor may comprise Dorsomorphin. In some cases, Dorsomorphin may inhibit ALK2, ALK3, ALK6, or a combination thereof. In some cases, a BMP receptor/AMPK inhibitor does not specifically inhibit ALK5.

3. Retinoic acid receptor (RAR) agonists

A chemical reprogramming factor may comprise an activator or agonist of RAR. RAR comprises a nuclear receptor transcription factor. RAR may comprise RAR-α or RAR-β. RAR may be activated by retinoic acids. The retinoic acid may comprise all-trans retinoic acid or 9-cis retinoic acid. RAR may form heterodimer with RXR (le Marie et al. 2019; Cells . 2019 Nov 5; 8 (11) : 1392., which is herein incorporated by reference in its entirety) . The RAR/RXR dimer with corepressor protein may inhibit the transcription from retinoic acid response elements (RAREs) (le Marie 2019) . Binding of RAR agonist to RAR can lead to a dissociation of the corepressor protein from the RAR/RXR complex. It can also lead to binding of RAR/RXR complex with the coactivator protein. Such binding can facilitate transcription of genes comprising the RAREs (le Marie 2019) .

An RAR agonist may comprise TTNPB ( [4- [ (E) -2- (5, 6, 7, 8-Tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthalenyl) -1-propenyl] benzoic acid] ) ; Ch 55 ( [4- [ (1E) -3- [3, 5-bis (1, 1-Dimethylethyl) phenyl] -3-oxo-1-propenyl] benzoic acid] ) ; or AM580 ( [4- [ (5, 6, 7, 8-Tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthalenyl) carboxamido] benzoic acid] ) ; or any combination thereof. An RAR agonist may comprise TTNPB. An RAR agonist may comprise Ch 55. Ch 55 may be a synthetic retinoid that has high affinity for RAR-α and RAR-β receptors and low affinity for cellular retinoic acid binding protein (CRABP) . An RAR agonist may comprise AM580. AM580 may be an analog of retinoic acid that acts as a selective RARα agonist.

4. S-adenosyl-L-homocysteine (SAH) hydrolase inhibitors

A chemical reprogramming factor may comprise a chemical compound that can inhibit SAH hydrolase. SAH hydrolase may catalyze reversible hydration of SAH into adenosine and homocysteine (Xiao et al. 2019; Circulation . 2019 May 7; 139 (19) : 2260-2277., which is herein incorporated by reference in its entirety) . SAH hydrolase may require NAD⁺ cofactor when catalyzing the hydration of SAH. Inhibition of SAH hydrolase can lead to accumulation of SAH, which in turn inhibits methyltransferase that utilizes SAM (or S-Adenosyl methionine) as the methyl group donor (Xiao 2019) . Accordingly, inhibition of SAH hydrolase can lead to inhibition of methylation of various cellular factors (Xiao 2019) .

A SAH hydrolase inhibitor may comprise 3-deazaneplanocin A (DZNep, [ (1S, 2R, 5R) -5- (4-Amino-1H-imidazo [4, 5-c] pyridin-1-yl) -3- (hydroxymethyl) -3-cyclopentene-1, 2-diol] ) ; (-) Neplanocin A (NepA, [5R- (6-amino-9H-purin-9-yl) -3- (hydroxymethyl) -3-cyclopentene-1S, 2R-diol] ) ; Adenozine periodate oxidized (Adox, [ (2S) -2- [ (1R) -1- (6-aminopurin-9-yl) -2-oxoethoxy] -3-hydroxypropanal] ) ; or 3-deazaadenosine (DZA, [1-β-D-ribofuranosyl-1H-imidazo [4, 5-c] pyridin-4-amine] ) or any combination thereof. A SAH hydrolase inhibitor may comprise DZNep. A SAH hydrolase inhibitor may comprise NepA. A SAH hydrolase inhibitor may comprise Adox. A SAH hydrolase inhibitor may comprise DZA.

5. Disruptor of telomeric silencing 1-like (Dot1L) inhibitors

A chemical reprogramming factor may comprise a Dot1L inhibitor. Dot1L is a histone H3 lysine 79 (H3K79) methyltransferase. Dot1L can catalyze the methylation of H3K79 (Kari et al. 2019; Clin Epigenetics . 2019 Jan 7; 11 (1) : 4., which is herein incorporated by reference in its entirety) . Dot1L is a non-SET domain containing methyltransferase known to catalyze mono-, di-, and tri-methylation of H3K79. Inhibition of Dot1L may decrease the methylation of H3K79 (Kari 2019) . Inhibition of Dot1L may decrease the phosphorylation of H2AX at serine 139 by specific DNA damage response-associated members of the phosphatidylinositol-3-kinase family induced by DNA damages (Kari 2019) . Inhibition of Dot1L may homologous recombination-mediated double strand break repairs (Kari 2019) .

A Dot1L inhibitor may comprise SGC 0946 ( [1- [3- [ [ [ (2R, 3S, 4R, 5R) -5- (4-Amino-5-bromo-7H-pyrrolo [2, 3-d] pyrimidin-7-yl) -3, 4-dihydroxytetrahydrofuran-2-yl] methyl] (isopropyl) amino] propyl] -3- [4- (2, 2-dimethylethyl) phenyl] urea] ) ; EPZ004777 ( [7- [5-Deoxy-5- [ [3- [ [ [ [4- (1, 1-dimethylethyl) phenyl] amino] carbonyl] amino] propyl] (1-methylethyl) amino] -β-D-ribofuranosyl] -7H-pyrrolo [2, 3-d] pyrimidin-4-amine] ) ; or EPZ5676 [ (2R, 3R, 4S, 5R) -2- (6-amino-9H-purin-9-yl) -5- ( ( ( (1r, 3S) -3- (2- (5- (tert-butyl) -1H-benzo [d] imidazol-2-yl) ethyl) cyclobutyl) (isopropyl) amino) methyl) tetrahydrofuran-3, 4-diol] ) ; or any combination thereof. A Dot1L inhibitor may comprise SGC 0946. A Dot1L inhibitor may comprise EPZ5676. A Dot1L inhibitor may comprise EPZ004777.

6. Histone deacetylase inhibitors

A chemical reprogramming factor may comprise a histone deacetylase inhibitor. Histone deacetylase may catalyze the deacetylation of histones. For example, a histone deacetylase may remove acetyl groups from an ε-N-acetyl lysine amino acid of a histone. Histone deacetylases may comprise class I, class IIA, class IIb, class III, and class IV histone deacetylase (Seto et al 2014; Cold Spring Harb Perspect Biol . 2014 Apr 1; 6 (4) : a018713., which is herein incorporated by reference in its entirety) . A class I histone deacetylase may comprise HDAC1, HDAC2, HDAC3, or HDAC8. A class IIA histone deacetylase may comprise HDAC4, HDAC5, HDAC7, or HDAC9. A class IIB histone deacetylase may comprise HDAC6 or HDAC10. A class IIII histone deacetylase may comprise sirtuin family (SIRT1, SIRT2, SIRT3, SIRT4, SIRT6, and/or SIRT7) or Sir2 (in budding yeast) . A class IV histone deacetylase may comprise HDAC11. A histone deacetylase inhibitor may inhibitor any one or any combinations of histone deacetylase described herein. Inhibition of a histone deacetylase may increase acetylation of histone (Seto 2014) . Inhibition of a histone deacetylase may also induce cell growth arrest, cell differentiation, or apoptosis (Seto 2014) . Inhibition of a histone deacetylase may also inhibit cancer cell growth (Seto 2014) .

A histone deacetylase inhibitor may comprise valproic acid (VPA) ; apicidin ( [cyclo (N-O-methyl-L-tryptophanyl-L-isoleucinyl-D-pipecolinyl -L-2-amino-8-oxodecanoyl) ] ; LMK235 ( [N- [ [6- (Hydroxyamino) -6-oxohexyl] oxy] -3, 5-dimethylbenzamide] ) ; MS275 ( [ (Pyridin-3-yl) methyl 4- (2-aminophenylcarbamoyl) benzylcarbamate] ) ; CI 994 ( [N-acetyldinaline4- (Acetylamino) -N- (2-aminophenyl) benzamide] ) ; Depsipeptide; KD 5170 ( [S- [2- [6- [ [ [4- [3- (Dimethylamino) propoxy] phenyl] sulfonyl] amino] -3-pyridinyl] -2-oxoethyl] ethanethioc acid ester] ; sodium, 4-pehynl butyrate; sodium butyrate ( [Butanoic acid sodium salt] ) ; or UF 010 ( [4-Bromo-N'-butylbenzohydrazide) ) ; HDACi IV; or any combination thereof. A histone deacetylase inhibitor may comprise VPA. A histone deacetylase inhibitor may comprise LMK235. A histone deacetylase inhibitor may comprise MS275. A histone deacetylase inhibitor may comprise CI 994. A histone deacetylase inhibitor may comprise KD5170. A histone deacetylase inhibitor may comprise Depsipeptide. A histone deacetylase inhibitor may comprise sodium butyrate. A histone deacetylase inhibitor may comprise UF 010.

7. B-Raf inhibitors

A chemical reprogramming factor may comprise a B-Raf inhibitor. B-Raf may comprise a protooncogene. B-Raf may also be referred to as B-Raf or v-Raf murine sarcoma viral oncogene homolog B. B-Raf is a serine/threonine-protein kinase. Activated RAS or RAS-GTP may activate B-Raf (Olsen et al. 2020; Sci Rep . 2020 Nov 18; 10 (1) : 20113; which is herein incorporated by reference in its entirety) . Activation of B-Raf may lead to increased phosphorylation of MEK, which in turn lead to increased phosphorylation of ERK (Olsen 2020) . Activation of B-Raf may also lead increased cell proliferation (Olsen 2020) . Inhibition of B-Raf may lead to decreased phosphorylation of MEK or ERK. Activation of B-Raf may lead decreased cell proliferation (Olsen 2020) .

A B-Raf inhibitor may comprise SB590885 ( [5- [2- [4- [2- (Dimethylamino) ethoxy] phenyl] -5- (4-pyridinyl) -1H-imidazol-4-yl] -2, 3-dihydro-1H-inden-1-one oxime] ) ; Vemurafenib; RAF265 (CHIR-265) (Selleckhchem catalog No. S2161) ; or PLX4720 (Selleckhchem catalog No. S11525) ; or any combination thereof. A B-Raf inhibitor may comprise SB590885. SB590885 may be a potent B-Raf inhibitor with an inhibitor constant (Ki) of 0.16 nM in a cell-free assay. SB590885 may be 11-fold greater selectivity for B-Raf over c-Raf. SB590885 may not inhibit other human kinases. A B-Raf inhibitor may comprise Vemurafenib. A B-Raf inhibitor may comprise RAF265. A B-Raf inhibitor may comprise PLX4720.

8. Wnt inhibitors

A chemical reprogramming factor may comprise a Wnt inhibitor (or a Wnt signaling inhibitor) . Wnt signaling pathway can be regulated by binding of a Wnt-protein ligand to a Frizzled family receptor. In canonical Wnt signaling pathway, binding of the Wnt ligand to the Frizzled family receptor can lead to a stabilization of beta-catenin, which in turn binds to Transcription factors of the T-cell family (TCF) and increases the transcription of genes under the regulation of Wnt-regulated enhancer. Wnt-signaling pathway may also comprise noncanonical planar cell polarity pathway and noncanonical Wnt/calcium pathway (Ramakrishnan et al., F1000Res . 2017 May 24; 6: 746., which is herein incorporated by reference in its entirety) . Inhibition of Wnt may lead to a decreased expression of beta-catenin. Activation or inhibition of Wnt signaling pathway can be assayed using TOP-flash assay, described in Molenaar et al., 1996; Cell . 1996 Aug 9; 86 (3) : 391-9, which is herein incorporated by reference in its entirety) . Inhibition of Wnt signaling may also lead to increased expression of SPATS1 gene (see, for example, Zhai et al, Cellular Signalling. 22 (11) : 1753–60, which is herein incorporated by reference in its entirety) .

A Wnt inhibitor may comprise IWP-2 ( [N- (6-methyl-2-benzothiazolyl) -2- [ (3, 4, 6, 7-tetrahydro-4-oxo-3-phenylthieno [3, 2-d] pyrimidin-2-yl) thio] -acetamide] ) ; WNT-C59 ( [4- (2-Methyl-4-pyridinyl) -N- [4- (3-pyridinyl) phenyl] benzeneacetamide] ) ; XAV-939 ( [3, 5, 7, 8-Tetrahydro-2- [4- (trifluoromethyl) phenyl] -4H-thiopyrano [4, 3-d] pyrimidin-4-one] ) ; or IWR-1 (Selleckchem catalog No. S7086) ; or any combination thereof. A Wnt inhibitor may comprise IWP-2. A Wnt inhibitor may comprise WNT-C59. A Wnt inhibitor may comprise XAV-939. A Wnt inhibitor may comprise IWR-1.

9. Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitors

A chemical reprogramming factor may comprise a ROCK inhibitor. ROCK is a kinase of family of serine-threonine specific protein kinases. ROCK may comprise ROCK1 or ROCK2. In some cases, ROCK can have a kinase domain, a coiled-coil region and a Pleckstrin homology (PH) domain (Tonges et al, 2011, Front Mol Neurosci. 2011; 4: 39., which is herein incorporated by reference in its entirety) . ROCK may be a effector of RhoA-GTP. Binding of Rho-A may alleviate the autoinhibition by the PH domain to its kinase activity (Tonges 2011) . Inhibition of ROCK may decrease phosphorylation of MLC (Tonges 2011) . Inhibition of ROCK may increase the activity of MLC phosphatase (Tonges 2011) . Inhibition of ROCK of decrease the activity of LIMK, which can lead to decreased phosphorylation of cofilin and increases actin depolymerization, and vice versa (Tonges 2011) .

A ROCK inhibitor may comprise Y27632 ( [ (+) - (R) -trans-4- (1-aminoethyl) -N- (4-pyridyl) cyclohexanecarboxamide+++ dihydrochloride) ] ) ; or Tzv (thiazovivin) ; or any combination thereof. A ROCK inhibitor may comprise Y27632. A ROCK inhibitor may comprise Tzv.

10. CBP/p300 bromodomain inhibitors

A chemical reprogramming factor may comprise a CBP/p300 bromodomain inhibitor. transcription coactivators CREB binding protein (CBP) and p300 are transcriptional coactivators. CBP and p300 may be acetyltransferases that mediate histone 3 lysine 27 acetylation (H3K27ac) . In some cases, CBP and p300 have redundant functions (Martire et al 2020; BMC Mol Cell Biol . 2020 Jul 20; 21 (1) : 55., which is herein incorporated by reference in its entirety) . Inhibition of CBP or p300 may lead to a decreased acetylation of H3K27 or H3K18 (Raisner et al., 2018; Cell Rep . 2018 Aug 14; 24 (7) : 1722-1729., which is herein incorporated by reference in its entirety) . Inhibition of CBP and/or p300 may lead to decreased expression of target genes, such as Myc and others (Raiser 2018) .

A CBP/p300 bromodomain inhibitor may decrease the acetyltransferase activity of CBP and/or p300. A CBP/p300 bromodomain inhibitor may comprise SGC-CBP30 ( [2- [2- (3-chloro-4-methoxyphenyl) ethyl] -5- (3, 5-dimethyl-4-isoxazolyl) -1- [ (2S) -2- (4-morpholinyl) propyl] -1H-benzimidazole] ) ; I-CBP112; GNE272; or GNE409; or any combination thereof. A CBP/p300 bromodomain inhibitor may comprise SGC-CBP30. A CBP/p300 bromodomain inhibitor may comprise I-CBP112. A CBP/p300 bromodomain inhibitor may comprise GNE272. A CBP/p300 bromodomain inhibitor may comprise GNE409.

11. Menin-MLL interaction inhibitors

A chemical reprogramming factor may comprise a Menin-MLL interaction inhibitor. Menin may be encoded by MEN1 in human. Menin may be a tumor suppressor (Cierpicki et al., 2014 Future Med Chem. 2014 Mar; 6 (4) : 447-62.; which is herein incorporated by reference in its entirety) . MLL may comprise a histone methyltransferase of the trithorax family (Cierpicki 2014) . MLL may comprise MLL1 or MLL2. Binding of Menin and MLL may increase the transcriptional activity of MLL. Inhibition of Menin, MLL, and or Menin-MLL interactions may lead to decreased expressions of Hoxa9, Hoxc6, and/or Hoxc8 (Cierpicki 2014) . Inhibition of Menin, MLL, and or Menin-MLL interactions may also decrease the expression of p27 and p18 cyclin-dependent kinase (CDK) inhibitors (Melne et al., 2005; Proc Natl Acad Sci U S A . 2005 Jan 18; 102 (3) : 749-54, which is herein incorporated by reference in its entirety) .

A Menin-MLL interaction inhibitor may comprise VTP50469, MI3454, or WDR5-IN-4, or any combination thereof. A Menin-MLL interaction inhibitor may comprise VTP50469. A Menin-MLL interaction inhibitor may comprise MI3454. A Menin-MLL interaction inhibitor may comprise WDR5-IN-4.

12. G protein-coupled receptor Smoothened agonists

A chemical reprogramming factor may comprise a G protein-coupled receptor Smoothened agonist. Smoothened may be encoded by the SMO gene (Arensdorf et al., 2016; Trends Pharmacol Sci . 2016 Jan; 37 (1) : 62-72., which is herein incorporated by reference in its entirety) . Smoothened may be a class F G protein-coupled receptor. Smoothened is a component of Hedgehog signaling pathway. Without the ligand Hedgehog, receptor Patched inhibits Smoothened. Once Hedgehog binds to Patched, Patched is internalized and degraded, activating Smoothened. Activated Smoothened can lead to the activation of Gli family transcription factors, which leads to increased expression of Ptch1, Ptch2, Gli1, CCND2, CCNE1, MYCN, BCL2, ABCG2, FGF4, VEGFA, PAX6, PAX7, PAX9, JAG1, or FOXM1 (Skoda et al., 2018; Bosn J Basic Med Sci . 2018 Feb 20; 18 (1) : 8-20., which is herein incorporated by reference in its entirety) .

A G protein-coupled receptor Smoothened agonist may comprise SAG [3-chloro-N- [4- (methylamino) cyclohexyl] -N- [ (3-pyridin-4-ylphenyl) methyl] -1-benzothiophene-2-carboxamide; hydrochloride, ] ) ; Purmorphamine; Hg-Ag1.5; or Sonic Hedgehog protein (Shh) , or any combination thereof. A G protein-coupled receptor Smoothened agonist may comprise SGA. A G protein-coupled receptor Smoothened agonist may comprise Purmorphamine. A G protein-coupled receptor Smoothened agonist may comprise Hg-Ag1.5. A G protein-coupled receptor Smoothened agonist may comprise Shh. Shh can be a human Shh. Shh can also be a mammalian Shh.

13. JAK1/2 (Jak1/2) inhibitors

A chemical reprogramming factor may comprise a Jak1/2 inhibitor. Jak may comprise Janus kinase. Jak1/2 may comprise tyrosine kinases. Jak1/2 may regulate cytokine signaling via interaction with type I/II cytokine receptors. These cytokine receptors can comprise interferon receptors, GM-CSF receptors, the gp130 receptors, single chain receptors, IL-2 receptors, IL-4 receptors, CT-1R receptors, CNTF receptors, NNT-1 receptors, Leptin receptors (Brooks et al., 2014; Science . 2014 May 16; 344 (6185) : 1249783. and Gadina et al., 2001; Curr Opin Immunol . 2001 Jun; 13 (3) : 363-73, each of which is herein incorporated by reference in its entirety) . Inhibitions of Jak1/2 may lead to decreased expressions of decreased phosphorylation of STAT, which leads to decreased expressions of target genes, such as Fas, Bcl-2, or Bcl-X. Inhibitions of Jak1/2 may lead to increased apoptosis Hu et al., 2021; Signal Transduct Target Ther . 2021 Nov 26; 6 (1) : 402., which is herein incorporated by reference in its entirety) .

A Jak1/2 inhibitor can comprise Ruxolitinib ( [ (3R) -3-cyclopentyl-3- [4- (7H-pyrrolo [2, 3-d] pyrimidin-4-yl) pyrazol-1-yl] propanenitrile] ) ; Tofacitinib; AZD1480; Baricitinib; S-Ruxolitinib; or Fedratinib; or any combination thereof. A Jak1/2 inhibitor can comprise Ruxolitinib. A Jak1/2 inhibitor can comprise Tofacitinib. A Jak1/2 inhibitor can comprise AZD1480. A Jak1/2 inhibitor can comprise Baricitinib. A Jak1/2 inhibitor can comprise S-Ruxolitinib. A Jak1/2 inhibitor can comprise Fedratinib.

14. Serine-threonine kinase Akt inhibitors

A chemical reprogramming factor may comprise an Akt inhibitor. Akt activation may activate mTOR via the phosphorylation of mTOR at Ser2448 and lead to the activation of complex mTORC1 and/or inhibit TSC1/TSC2. Activated mTORC1 may increase the phosphorylation of 70S6K1 or eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1) . Phosphorylation of 4EBP-1 can promote the expression of hypoxia-inducible factor 1α, cyclin D1, and/or c-Myc, which leads to angiogenesis or cell cycle progression. Akt activation may lead to increased cell proliferation. Inhibition of Akt can decrease the phosphorylation of mTOR, TSC1/2, 4eBP-1, or 70S6K1. Inhibition of Akt can decrease the expression of hypoxia-inducible factor 1α, cyclin D1, and/or c-Myc (Li et al., 2020; Cell Death Dis . 2020 Sep 24; 11 (9) : 797.; which is herein incorporated by reference in its entirety) .

An Akt inhibitor may comprise AKT Kinase Inhibitor (AKTi; [3- [2- (4-amino-1, 2, 5-oxadiazol-3-yl) -7- (3-aminopropoxy) -1-ethylimidazo [4, 5-c] pyridin-4-yl] prop-2-yn-1-ol] ) .

15. c-Jun kinase inhibitors

A chemical reprogramming factor may comprise a c-Jun kinase inhibitor. c-Jun kinase is a MAP kinase family kinase. c-Jun kinase may phosphorylate c-Jun on Ser-63 and Ser-73 within its transcriptional activation domain. Activation of c-Jun kinase may lead to the activation of AP1 target genes including FosB, WEE1, PVR, MAP1LC3B and/or LGALS3) , and vice versa (Schummer et al., 2016; Cancer Biol Ther . 2016 May 3; 17 (5) : 486-97, which is herein incorporated by reference in its entirety.

A c-Jun kinase inhibitor may comprise JNKIN8 ( [3- [ [ (E) -4- (dimethylamino) but-2-enoyl] amino] -N- [3-methyl-4- [ (4-pyridin-3-ylpyrimidin-2-yl) amino] phenyl] benzamide] ) ; JNKIN7; JNKIN5; or JNKIN12; or any combination thereof. A c-Jun kinase inhibitor may comprise JNKIN8. A c-Jun kinase inhibitor may comprise JNKIN5. A c-Jun kinase inhibitor may comprise JNKIN12. A c-Jun kinase inhibitor may comprise JNKIN7.

16. p38 MAPK inhibitors

A chemical reprogramming factor may comprise a p38 MAPK inhibitor. As used herein, a p38 MAPK (or p38 mitogen-activated protein kinase) is a kinase of class of mitogen-activated protein kinases. p38 MPAK may comprise p38-α; p38-β; p38-γ; or p38-δ. Activation of p38 MPAK may increase the phosphorylation of MAPKAP kinase 2, ATF2, Mac, MEF2, or p53, and vice versa (Rawas et al., 2020; Int J Mol Sci . 2020 Jul 8; 21 (14) : 4833, which is herein incorporated by reference in its entirety) . Inhibition of p38 MPAK may decrease the expression of Jun, Fos, Myc, Egr-1, Maff, Sox2, Runx2, or others described in Whitmarsh 2010; BMC Biol . 2010 Apr 27; 8: 47, which is herein incorporated by reference in its entirety) . Inhibition of p38 MPAK may decrease cell proliferation.

A p38 MAPK inhibitor may comprise BIRB796 ( [1- [5-tert-butyl-2- (4-methylphenyl) pyrazol-3-yl] -3- [4- (2-morpholin-4-ylethoxy) naphthalen-1-yl] urea] ) ; SB203580; or SB202190; or any combination thereof. A p38 MAPK inhibitor may comprise BIRB796. A p38 MAPK inhibitor may comprise SB2033580. A p38 MAPK inhibitor may comprise SB202190.

17. MEK inhibitors

A chemical reprogramming factor may comprise a MEK inhibitor. A MEK (mitogen-activated protein kinase kinase) is also known as MAP2K or MAPKK, may be a dual-specificity kinase enzyme which phosphorylates mitogen-activated protein kinase (MAPK) . There can be seven genes that encode various MEKs, such as MAP2K1 (encoding MEK1) , MAP2K2 (encoding MEK1) , MAP2K3 (encoding MEK3) , MAP2K4 (encoding MEK4) , MAP2K5 (encoding MEK5) , MAP2K6 (encoding MEK6) , and MAP2K7 (encoding MEK7) . MEKs may activate p38 MAPK (e.g., by MKK3 and MKK6) , JNK (e.g., by MKK4 and MKK7) , and ERK (e.g., by MEK1 and MEK2) (Dérijard B, et al. (1995) Science. 267 (5198) : 682–5, which is herein incorporated by reference in its entirety) . MEK may phosphorylate and activate a MAPK (e.g., a p38 MAPK) . In some case, the inhibition of MEK and the inhibition of MAPK may lead to similar changes of biological activities. A MEK inhibitor described herein is a direct inhibitor of a MEK protein, and does not directly inhibit a MAPK protein (e.g., does not inhibit a MAPK protein in an in vitro cell-free kinase assay study that does not involve MEK proteins) .

A MEK inhibitor may also comprise PD0325901, AZD8330, or TAK-733, or any combination thereof. A MEK inhibitor may comprise PD0325901. A MEK inhibitor may comprise AZD8330. A MEK inhibitor may comprise TAK-733.

18. Adenosine kinase inhibitors

A chemical reprogramming factor may comprise an adenosine kinase inhibitor. An adenosine kinase may phosphorylate adenosine to adenosine monophosphate using the gamma phosphate of ATP. Inhibition of adenosine kinase may lead to an increase amount of SAH. Increased amounts of SAH may inhibit transmethylation reactions (Fox et al, 1978; Annu Rev Biochem . 1978; 47: 655-86., which is herein incorporated by reference in its entirety) .

An adenosine kinase inhibitor may comprise 5-Iodotubercidin (5-ITU; [ (2R, 3R, 4S, 5R) -2- (4-amino-5-iodopyrrolo [2, 3-d] pyrimidin-7-yl) -5- (hydroxymethyl) oxolane-3, 4-diol] ) ; or ABT 702 dihydrochloride; or any combination thereof . An adenosine kinase inhibitor may comprise 5-ITU. An adenosine kinase inhibitor may comprise ABT 702) .

19. SETD2 inhibitors

A chemical reprogramming factor may comprise a SETD2 inhibitor. SETD2 may comprise a histone methyltransferase that methylates the lysine 36 of histone H3 (H3K36) . SETD2 may mediates mono-, di-, or tri-methylation of H3. Inhibition of SETD2 may decrease the phosphorylation of H3 at K36. In some cases, inhibition of SETD2 may increase the frequency of deletion mutations induced by microhomology-mediated end joining (Pfister et al., 2014; Cell Rep . 2014 Jun 26; 7 (6) : 2006-18., which is herein incorporated by reference in its entirety) . Inhibition of SETD2 may increase the frequency of DNA repair by microhomology-mediated end joining. Inhibition of SETD2 may decrease homologous recombination repair.

A SETD2 inhibitor may comprise SETD2-IN-1 ( [N- [ (1R, 3S) -3- (4-acetylpiperazin-1-yl) cyclohexyl] -4-fluoro-7-methyl-1H-indole-2-carboxamide; 2, 2, 2-trifluoroacetic acid] ) ; EPZ-719; or MMSET-IN-1; or any combination thereof. A SETD2 inhibitor may comprise SETD2-IN-1. A SETD2 inhibitor may comprise EPZ-719. A SETD2 inhibitor may comprise MMSET-IN-1.

20. casein kinase 2 inhibitors

A chemical reprogramming factor may comprise a casein kinase 2 inhibitor. Casein kinase 2 (CK2) may be a serine/threonine protein kinase. CK2 may use ATP or CTP as phosphate sources. CK2 may phosphorylate AKT, STAT, beta-catenin, or androgen receptors (Borgo et al., 2021; Signal Transduct Target Ther . 2021 May 17; 6 (1) : 183; Signal Transduct Target Ther . 2021 May 17; 6 (1) : 183) . Inhibition of CK2 may lead to decreased phosphorylation of AKT, STAT, beta-catenin, or androgen receptors. Inhibition of CK2 may lead to decreased expression of genes regulated by AKT beta-catenin as described elsewhere in this disclosure.

A casein kinase 2 inhibitor may comprise CX-4945 ( [5- (3-chloroanilino) benzo [c] [2, 6] naphthyridine-8-carboxylic acid] ) .

21. Inhibitors of histone demethylation

A chemical reprogramming factor may comprise an inhibitor of histone demethylation. Histone demethylation may lead to global activation of gene expression. Inhibition of histone demethylation may result in an increased level of histone methylation. An inhibitor of histone demethylation may comprise Tranylcypromine.

22. Adenosine kinase inhibitors

A chemical reprogramming factor may comprise an adenosine kinase inhibitor. Adenosine kinase is an enzyme that can catalyze the transfer of gamma-phosphate from

Adenosine triphosphate (ATP) to adenosine (ADO) leading to formation of Adenosine monophosphate (AMP) . Adenosine (ADO) is an endogenous protective regulator that can restore cellular energy balance in response to tissue trauma. Extracellular ADO can have a half-life of the order of seconds thus restricting its actions to tissues and cellular sites where it is released. Adenosine kinase can be the rate-limiting enzyme controlling extracellular ADO concentrations. Inhibition of AK can effectively increase ADO extracellular concentrations at tissue sites where pathophysiological changes occur. (Jarvis MF. Pharmacol Res Perspect. 2019 Jul 22; 7 (4) : e00506. doi: 10.1002/prp2.506. PMID: 31367385; PMCID: PMC6646803. )

An adenosine kinase inhibitor may comprise 5-Iodotubercidin ( (2R, 3R, 4S, 5R) -2- (4-amino-5-iodopyrrolo [2, 3-d] pyrimidin-7-yl) -5- (hydroxymethyl) oxolane-3, 4-diol, or also known as 7-iodo-7-carba-adenosine) or ABT 702 (5- (3-bromophenyl) -7- (6-morpholin-4-ylpyridin-3-yl) pyrido [2, 3-d] pyrimidin-4-amine) . An adenosine kinase inhibitor may comprise 5-Iodotubercidin

23. G9a inhibitors

A chemical reprogramming factor may comprise a G9a inhibitor. G9a is a histone methyltransferase that can specifically mediate mono-and di-methylation of histone H3 lysine 9 (H3K9) . Lysine methylation of histone H3 by G9a is associated with transcriptional repression. (Rada M et al. Oncogene. 2017; 36: 922–32; and Olcina MM et al. Oncogene. 2016; 35: 793–9) .

A G9a inhibitor may comprise Unc0224 (7- [3- (dimethylamino) propoxy] -6-methoxy-2- (4-methyl-1, 4-diazepan-1-yl) -N- (1-methylpiperidin-4-yl) quinazolin-4-amine) , Unc0638 (2-cyclohexyl-6-methoxy-N- (1-propan-2-ylpiperidin-4-yl) -7- (3-pyrrolidin-1-ylpropoxy) quinazolin-4-amine) , Unc0642 (2- (4, 4-difluoropiperidin-1-yl) -6-methoxy-N- (1-propan-2-ylpiperidin-4-yl) -7- (3-pyrrolidin-1-ylpropoxy) quinazolin-4-amine) , or Bix01294 (N- (1-benzylpiperidin-4-yl) -6, 7-dimethoxy-2- (4-methyl-1, 4-diazepan-1-yl) quinazolin-4-amine) .

24. DNA methyltransferase (DNMT) inhibitors

A chemical reprogramming factor may comprise a DNMT inhibitor. In biochemistry, there is a family of DNA methyltransferases, which comprises a conserved set of DNA-modifying enzymes that can have a central role in epigenetic gene regulation. Canonical DNMT enzymes can include DNMT1, DNMT3A and DNMT3B. (Lyko, F. Nat Rev Genet 19, 81–92 (2018) ) .

A DNMT inhibitor may comprise 5-Azacytidine (4-amino-1- [ (2R, 3R, 4S, 5R) -3, 4-dihydroxy-5- (hydroxymethyl) oxolan-2-yl] -1, 3, 5-triazin-2-one) , Decitabine (4-amino-1- [ (2R, 4S, 5R) -4-hydroxy-5- (hydroxymethyl) oxolan-2-yl] -1, 3, 5-triazin-2-one, also known as 5-Aza-2'-deoxycytidine or 2'-Deoxy-5-azacytidine) , RG108 ( (2S) -2- (1, 3-dioxoisoindol-2-yl) -3- (1H-indol-3-yl) propanoic acid) , or SGI-1027 (N- [4- [ (2-amino-6-methylpyrimidin-4-yl) amino] phenyl] -4- (quinolin-4-ylamino) benzamide) . A DNMT inhibitor may comprise 5-Azacytidine.

CELL CULTURE

In some cases, a composition may comprise a culture medium. Methods provided herein may comprise culturing a population of cells. Culturing a population of cells may comprise contacting the population of cells with a culture medium. Culturing a population of cells may comprise incubating the population of cells with the culture medium for a period of time. During or subsequent to the culturing, at least a subset of the population of cells may undergo cell proliferation. The subset of the population of cells may increase in cell numbers and/or cell mass. In some cases, during or subsequent to the culturing, a subset of the population of cells may give rise to a progeny or progenies.

During a conversion of a cell population, at least a subset of the population of cell may be converted to a different cell type. In some case, the subset of the population of cells may comprise at least a progeny of the subset of the population of cells.

The culture medium may comprise a chemical reprogramming factor. In some cases, the culture medium may not comprise A chemical reprogramming factor. In some cases, the culture medium may comprise at least a molecule for supporting the viability of a cell. In some cases, the culture medium may comprise at least a molecule for supporting the proliferation of a cell. A culture medium may comprise at least a molecule for supporting growth of a cell in vitro or ex vivo. A culture medium may comprise a peptide, a polypeptide, a growth factor, a carbon source, a nitrogen source, a mineral source, a vitamin source, water, salt, oxygen, or carbon dioxide, or any combination thereof.

The stage 1 methods provided herein may comprise plating a first population of cells or somatic cells onto a cell growth substrate. The cell growth substrate may comprise a culture plate. The culture plate may have a growth area of about 0.1 square centimeter (cm^2) , 0.2 cm^2, 0.3 cm^2, 0.4 cm^2, 0.5 cm^2, 0.6 cm^2, 0.7 cm^2, 0.8 cm^2, 0.9 cm^2, 1 cm^2, 1.1 cm^2, 1.2 cm^2, 1.3 cm^2, 1.4 cm^2, 1.5 cm^2, 1.6 cm^2, 1.7 cm^2, 1.8 cm^2, 1.9 cm^2, 2 cm^2, 2.1 cm^2, 2.2 cm^2, 2.3 cm^2, 2.4 cm^2, 2.5 cm^2, 2.6 cm^2, 2.7 cm^2, 2.8 cm^2, 2.9 cm^2, 3 cm^2, 3.1 cm^2, 3.2 cm^2, 3.3 cm^2, 3.4 cm^2, 3.5 cm^2, 3.6 cm^2, 3.7 cm^2, 3.8 cm^2, 3.9 cm^2, 4 cm^2, 4.1 cm^2, 4.2 cm^2, 4.3 cm^2, 4.4 cm^2, 4.5 cm^2, 4.6 cm^2, 4.7 cm^2, 4.8 cm^2, 4.9 cm^2, 5 cm^2, 6 cm^2, 7 cm^2, 8 cm^2, 9 cm^2, 10 cm^2, 20 cm^2, 30 cm^2, 40 cm^2, 50 cm^2 or more.

The first population of stage 1 cells or somatic cells may be plated on the cell growth substrate at a density. Such a density may comprise be at least about 1x10^3 cells per cm^2 cell growth area, 2.5x10^3 cells per cm^2 cell growth area, 5x10^3 cells per cm^2 cell growth area, 1x10^4 cells per cm^2 cell growth area, 2.5x10^4 cells per cm^2 cell growth area, 5x10^4 cells per cm^2 cell growth area, 1x10^5 cells per cm^2 cell growth area, 2.5x10^5 cells per cm^2 cell growth area, 5x10^5 cells per cm^2 cell growth area, 1x10^6 cells per cm^2 cell growth area, 2.5x10^6 cells per cm^2 cell growth area, 5x10^6 cells per cm^2 cell growth area, 1x10^7 cells per cm^2 cell growth area, 2.5x10^7 cells per cm^2 cell growth area, 5x10^7 cells per cm^2 cell growth area, 1x10^8 cells per cm^2 cell growth area, 2.5x10^8 cells per cm^2 cell growth area, 5x10^8 cells per cm^2 cell growth area or more. Such a density may comprise be at most about 1x10^3 cells per cm^2 cell growth area, 2.5x10^3 cells per cm^2 cell growth area, 5x10^3 cells per cm^2 cell growth area, 1x10^4 cells per cm^2 cell growth area, 2.5x10^4 cells per cm^2 cell growth area, 5x10^4 cells per cm^2 cell growth area, 1x10^5 cells per cm^2 cell growth area, 2.5x10^5 cells per cm^2 cell growth area, 5x10^5 cells per cm^2 cell growth area, 1x10^6 cells per cm^2 cell growth area, 2.5x10^6 cells per cm^2 cell growth area, 5x10^6 cells per cm^2 cell growth area, 1x10^7 cells per cm^2 cell growth area, 2.5x10^7 cells per cm^2 cell growth area, 5x10^7 cells per cm^2 cell growth area, 1x10^8 cells per cm^2 cell growth area, 2.5x10^8 cells per cm^2 cell growth area, 5x10^8 cells per cm^2 cell growth area or more.

When culturing the cells provided herein, the methods may comprise passaging the cells. In some cases, passaging the cells may comprise removing a first composition from a population of cells, splitting the populations of cells into a subset of the population of cells, and incubating the split population of cells with a second composition. A passage my comprise the process of passaging the cells once. The first and second compositions for passaging may be comprise the same chemical make-up. The second composition may be new or haven’ t contacted a cell prior to contacting the split population of cells. Hence, passaging may comprise replacing a culture medium with a new culture medium. In some cases, the split population of cells may be about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%or the unsplit population of cells. The methods described herein may comprise at most 2, 3, 4, 5, or 6 passages. The methods may comprise at most 5 passages. The methods may comprise at most 4 passages. The methods may comprise at most 3 passages.

EQUIVELANT AMOUNTS FOR SUBSTITUING CHEMICAL REPROGRAMMING FACTORS

In some cases, two different chemical reprogramming factors may substitute each other in any of the compositions or methods described herein when converting, contacting, or incubating cells. In some cases, two different chemical reprogramming factors may substitute each other if they increase or decrease a same biological activity. In some cases, two different chemical reprogramming factors may substitute each other if they inhibit a same enzyme, a same protein-protein binding domain, or a same molecular interaction, or any combination thereof. When a first chemical reprogramming factor substitute a second chemical reprogramming factor, an amount of the first chemical reprogramming factor may be used. Such an amount of the first chemical reprogramming factor may comprise an equivalent amount of the first chemical reprogramming factor.

To identify the equivalent amount of the first chemical reprogramming factor, one or more assays may be carried out. For example, the first and second chemical reprogramming factors may activate or inhibit a same target (e.g., an enzyme or a protein-protein binding interaction. The equivalent concentration of the first chemical reprogramming factor may be derived using a biological assay that assay the same target. A measurable value may be derived individual biological assay, each assaying the same target using the first or second chemical reprogramming factor. The equivalent amount of the first chemical reprogramming factor may be one that has a measurable value that is within 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%of that of the second chemical reprogramming factor. In other cases, the equivalent concentration of the first chemical reprogramming factor may be derived using the methods described herein to convert cells. For example, the methods may assay the number of converted cells generated from a starting population of cells using one of the first and second chemical reprogramming factors (i.e., each of the first and second chemical reprogramming factors is assayed for how many converted cells are generated from a substantially same number of the starting population of cells (e.g., the numbers of cells are within at least 90%with each assay) . The equivalent amount of the first chemical reprogramming factor may be one that has a number of converted cells that is within 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%of that of the second chemical reprogramming factor.

EFFECTIVE AMOUNTS OF CHEMICAL REPROGRAMMING FACTORS FOR CELL CONVERSION

The methods or compositions provided herein may comprise an effective amount of a chemical reprogramming factor for cell conversion. An effective amount of a chemical reprogramming factor for cell conversions may comprise the amount of the chemical reprogramming factor that can facilitate the conversion of one cell type to another cell type. An effective amount of a chemical reprogramming factor for cell conversions comprises the amount of the chemical reprogramming factor (e.g., any amounts described herein) relative to a number of cells (e.g., any numbers of cells in a cell population before contacting one composition described herein, such that the contacting may converts at least a cell type from the cell population to another cell type) .

In some cases, a chemical reprogramming factor may have an effective range for converting cells. The effective range may comprise a lower amount and a higher amount of the chemical reprogramming factors. Within the effective range, the number of converted cells generated from a starting population of cells, using the methods or compositions described herein, may be substantially the same. This substantially the same number of cells may mean that the numbers of converted cells generated from the lower and the higher amounts of the chemical reprogramming factors are within 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

GENETIC MODIFICATION

A cell provided herein, or a progeny thereof may comprise a genome modification. The first cell population of stage 1 may comprise one or more cells with the genetic modification. The second cell population of stage 1 may comprise one or more cells with the genetic modification. The first cell population of stage 2 may comprise one or more cells with the genetic modification. The second cell population of stage 2 may comprise one or more cells with the genetic modification. The cell with the genetic modification may comprise a somatic cell, intermediate plastic state cell, pluripotent stem cell, or a progeny thereof, or a combination thereof. The cell with the genetic modification may comprise a somatic cell or the progeny thereof. The cell with the genetic modification may comprise an intermediate plastic state cell or a progeny thereof. The cell with the genetic modification may comprise a pluripotent stem cell or a progeny thereof.

In the methods described herein, at least a subset of a population of cells may comprise a genetic modification. During the cell conversion and/or culturing a converted cell derived from the subset of the population of cells or any progenies thereof may comprise the same genetic modification.

A genetic modification of a cell may comprise a change in the genetic material of the cell. A genetic modification may comprise a change in the make-up of a genome of a cell. The genetic material may comprise a genome of a cell. The genetic material may comprise chromosomal DNA or extra-chromosomal DNA. The extra-chromosomal DNA may comprise a mitochondrial DNA. A genetic modification may comprise a mutation. The mutation may comprise a substitution, deletion, or insertion of the genome. In other cases, the mutation also comprises a DNA rearrangement of the chromosomal or mitochondrial DNA. In some cases, a genetic modification may comprise insertions of exogenous genetic materials into a cell. In other cases, a genetic modification may comprise insertions of exogenous genetic materials into a genome of a cell.

A genetic modification may comprise random mutagenesis of a genome of the cell. A genetic modification may comprise homologous or non-homologous recombination within the genome of a cell. A genetic modification may comprise utilizing a nuclease to generate the genetic modification. The nuclease may comprise an exonuclease, an endonuclease, or a combination thereof. The nuclease may comprise an exonuclease. The nuclease may comprise an endonuclease. The endonuclease may comprise zinc finger nuclease (ZFN) , transcription activator like effector nuclease (TALEN) , homing endonuclease (HE) , meganuclease, MegaTAL, a clustered regularly interspaced short palindromic repeats (CRISPR) -associated endonuclease, or a combination thereof. The genetic modification may also comprise a reverse transcriptase. In some embodiments, an endonuclease may introduce one or more single-stranded breaks (SSBs) and/or one or more double-stranded breaks (DSBs) .

A genetic modification may comprise a cell carrying an exogenous nucleic acid. The exogenous nucleic acid may comprise DNA or ribonucleic acid (RNA) . The exogenous nucleic acid may comprise a virus. The exogenous nucleic acid may not comprise a virus. The exogenous nucleic acid may comprise a viral-based vector, a non-viral-based vector, or a combination thereof. The exogenous nucleic acid may comprise a viral-based vector. The exogenous nucleic acid may comprise a non-viral-based vector. The exogenous nucleic acid may comprise a viral-based vector and non-viral-based vector.

The exogenous nucleic acid may comprise a sequence that encodes a polypeptide, a non-coding nucleic acid molecule, or a combination thereof. The exogenous nucleic acid may comprise a sequence that encodes a polypeptide. The exogenous nucleic acid may comprise a sequence that encodes non-coding nucleic acid molecule. The exogenous nucleic acid may comprise a sequence that encodes a polypeptide and a non-coding nucleic acid molecule. The non-coding nucleic acid molecule may comprise a micro ribonucleic acid (miRNA) , a long non-coding RNA (lncRNA) , a ribosomal ribonucleic acid (rRNA) , a silencing ribonucleic acid (siRNA) , a short hairpin RNA (shRNA) , a ribozyme, or any combinations thereof.

A cell comprising a genetic modification may have reduced immunogenicity. In some cases, the cell comprising a genetic modification may elicit reduced immune response of a subject against the cell, when the cell is administered to the subject, relative to a comparable cell that does not comprise the genetic modification. In some cases, the reduction of the immunogenicity of a cell comprising the genetic modification may be at least about 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%, relative to a comparable cell that does not comprise the genetic modification. In some cases, the reduction of the immunogenicity of a cell comprising the genetic modification may be at most about 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100%, relative to a comparable cell that does not comprise the genetic modification. When comparing immunogenicity, a comparable cell to a cell may comprise a cell that with the same cell type as the cell. The comparable cell may not comprise the same genetic modification of the cell comprising the genetic modification.

METHODS OF USES

Identification of a readily available source of stem cells, progenitor cells, dedifferentiated cells or cells with potency that can give rise to a desired cell type or morphology can be beneficial for therapeutic treatments or tissue engineering.

The cells obtained by methods of provided can comprise CiPSCs, intermediate plastic state cells, and epithelia-like cells that are readily available source of stem cells, progenitor cells, dedifferentiated cells, or cells with potency. In some cases, stem cells, progenitor cells, dedifferentiated cells, or cells with potency obtained by a method provided herein express at least one of stem cell related markers such as LIN28A, SALL4, OCT4 or NANOG. Intermediate plastic state cells or epithelia-like cells obtained by methods of this application may also be used similarly as source of stem cells, progenitor cells, dedifferentiated cells, or cells with potency. The availability of stem cells, progenitor cells, dedifferentiated cells or cells with potency and regenerative potentials can be beneficial in transplantation, tissue engineering, and regulation of angiogenesis, vasculogenesis, and cell replacement or cell therapies, as well as the prevention of certain diseases. Such stem cells or progenitor cells can also be used to introduce a gene into a subject as part of a gene therapy regimen. In addition, the cells are obtained by a method of this application comprising one or both of stages 1 or 2. For example, any of the pluripotent stem cells, epithelia-like cells, or intermediate plastic state cells may be directly induced to a desired cell type and implanted and delivered to the subject.

Providing differentiated somatic cells (re-differentiated cells)

Once established, a culture of stem cells may be used to produce progeny cells, for example, fibroblasts capable of producing new tissue. The CiPSCs (e.g., human CiPSCs or hCiPSCs) can be induced to differentiate into cells from any of the three germ layers, for example, skin and hair cells including epithelial cells, keratinocytes, melanocytes, adipocytes, cells forming bone, muscle and connective tissue such as myocytes, chondrocytes, osteocytes, alveolar cells, parenchymal cells such as hepatocytes, renal cells, adrenal cells, and islet cells, blood cells, retinal cells (and other cells involved in sensory perception, such as those that form hair cells in the ear or taste buds on the tongue) , and nervous tissue including nerves.

In one embodiment, the CiPSCs can be induced to differentiate into cells of ectodermal origin by exposing the cells to an "ectodermal differentiating" media. In another embodiment the CiPSCs can be induced to differentiate into cells of mesodermal origin by exposing the cells to "mesodermal differentiating media" . In still another embodiment, the CiPSCs can be induced to differentiate into cells of endodermal origin by exposing the cells to “endodermal media” . Components of “endodermal” , “mesodermal” and “ectodermal” media are known to one of skill in the art. Known cell surface markers can be used to verify that the cells are indeed differentiating into cells of the lineage of the corresponding cell culture medium. The most commonly accepted markers to confirm differentiation of the three germ layers are the expression of alpha fetal protein for endodermal cells, alpha smooth muscle actin for mesoderm, and Beta-III tubulin for ectoderm, all of which are normally expressed very early in the development of these tissues.

Differentiation of stem cells to fibroblasts or other cell types, followed by the production of tissue therefrom, can be triggered by specific exogenous growth factors or by changing the culture conditions (e.g., the density) of a stem cell culture. Methods for inducing differentiation of cells into a cell of a desired cell type can comprise, for example, CiPSCs can be induced to differentiate by adding a substance (e.g., a growth factor, enzyme, hormone, or other signaling molecule) to the cell's environment. The differentiated cells can be expanded in culture and stored for later retrieval and use.

The intermediate plasticity state cells, and epithelia-like cells are readily available source for generating to other cell types that can be triggered by specific exogenous growth factor, small molecules, over expression genes or by changing the culture conditions (e.g., the density) of a stem cell culture. The cells induced from the intermediate plasticity state cells, and epithelia-like cells can be different cell types including but not limited to somatic cells of hematological origin, skin derived cells, adipose cells, epithelial cells, endothelial cells, cells of mesenchymal origin, parenchymal cells (for example, hepatocytes, β cells) , neurological cells, and connective tissue cells.

1. Cell therapy

Therapeutic uses of the induced pluripotent stem cells can comprise transplanting the induced pluripotent stem cells, stem cell populations, or progeny thereof into individuals to treat a variety of pathological states including diseases and disorders resulting from cancers, wounds, neoplasms, injury, viral infections, diabetes, and the like. Treatment may entail the use of the cells to produce new tissue, and the use of the tissue thus produced. The cells may be implanted, injected, or otherwise administered directly to the site of tissue damage so that they will produce new tissue in vivo. In one embodiment, administration includes the administration of genetically modified CiPSCs or their progeny. In one embodiment, the CiPSCs are obtained from autologous cells i.e., the donor cells are autologous. However, the cells can be obtained from heterologous cells. In one embodiment, the donor cells are obtained from a donor genetically related to the recipient. In another embodiment, donor cells are obtained from a donor genetically un-related to the recipient.

If the CiPSCs are derived from a heterologous (non-autologous/allogenic) source compared to the recipient subject, concomitant immunosuppression therapy is typically administered, e.g., administration of the immunosuppressive agent cyclosporine or FK506. However, due to the immature state of the human induced pluripotent stem cells such immunosuppressive therapy may not be required. Accordingly, in one embodiment, the human induced pluripotent stem cells can be administered to a recipient in the absence of immunomodulatory (e.g., immunosuppressive) therapy. Alternatively, the cells can be encapsulated in a membrane, which permits exchange of fluids but prevents cell/cell contact. Transplantation of microencapsulated cells is described in Balladur et al., Surgery, 117: 189-94, 1995; and Dixit et al., Cell Transplantation 1: 275-79 (1992) , each of which is herein incorporated by reference in its entirety.

The CiPSCs can be induced to differentiate into cells from any of the three germ layers, for example, skin and hair cells including epithelial cells, keratinocytes, melanocytes, adipocytes, cells forming bone, muscle and connective tissue such as myocytes, chondrocytes, osteocytes, alveolar cells, parenchymal cells such as hepatocytes, renal cells, adrenal cells, and islet cells (e.g., alpha cells, delta cells, PP cells, and beta cells) , blood cells (e.g., leukocytes, erythrocytes, macrophages, and lymphocytes) , retinal cells (and other cells involved in sensory perception, such as those that form hair cells in the ear or taste buds on the tongue) , and nervous tissue including nerves.

(i) Diabetes

Diabetes mellitus (DM) is a group of metabolic diseases where the subject has high blood sugar, either because the pancreas does not produce enough insulin, or, because cells do not respond to insulin that is produced.

A replacement for insulin therapy is provision of islet cells to the patient in need of insulin. Shapiro et al., N Engl J Med., 343 (4) : 230-8 (2000) , which is herein incorporated by reference in its entirety, have demonstrated that transplantation of beta cells/islets provides therapy for patients with diabetes. Although numerous insulin types are commercially available, these formulations are provided as injectables. The human induced pluripotent stem cells can provide an alternative source of islet cells to prevent or treat diabetes. For example, induced pluripotent stem cells can be isolated and differentiated to a pancreatic cell type and delivered to a subject. Alternatively, the induced pluripotent stem cells can be delivered to the pancreas of the subject and differentiated to islet cells in vivo. Accordingly, the cells can be beneficial for transplantation in order to prevent or treat the occurrence of diabetes. Methods for reducing inflammation after cytokine exposure without affecting the viability and potency of pancreatic islet cells are disclosed for example in U.S. Patent No. 8,637,494 to Naziruddin, et al, which is herein incorporated by reference in its entirety.

(ii) Neurological disorders

Neurological disorders and conditions can be characterized by conditions involving the dysfunction and/or deterioration of neurons and/or glial cells, as a result of disease, hereditary conditions, or injury, such as traumatic or ischemic spinal cord or brain injury. Neurological disorders and conditions can comprise any disease or disorder or symptoms or causes or effects thereof involving dysfunction, damage, or deterioration of neurons and/or glial cells. Neurological disorders and conditions, to which the cells provided herein are applicable, can include, but are not limited to, neurodegenerative diseases and conditions.

The methods disclosed herein can comprise transplanting into a subject in need thereof NSCs, neural progenitors, neural precursors, or glial cells that have been expanded in vitro such that the cells can ameliorate the neurological disorders and conditions. Transplantation of the expanded cells (e.g., NSCs, neural progenitors, neural precursors, or glial cells) can be used to improve ambulatory function in a subject suffering from various forms of myelopathy with symptoms of spasticity, rigidity, seizures, paralysis, or any other hyperactivity of muscles. Methods for expanding and transplanting neural cells and neural progenitor cells for the treatment of different neurodegenerative conditions is disclosed for example, in U.S. Patent No. 8,236,299 to Johe, et. al, which is herein incorporated by reference in its entirety.

(iii) Cancer Therapy

Therapeutic uses of the CiPSCs and their progeny can comprise transplanting the induced pluripotent stem cells, stem cell populations, or progeny thereof into individuals to treat and/or ameliorate the symptoms associated with cancer. For example, in one embodiment, the CiPSCs can be administered to cancer patients who have undergone chemotherapy that has killed, reduced, or damaged cells of a subject. In a typical stem cell transplant for cancer, high doses of chemotherapy are used, often along with radiation therapy, to aim to destroy all the cancer cells. This treatment also kills the stem cells in the bone marrow. Soon after treatment, stem cells are given to replace those that were destroyed.

In some cases, the CiPSCs can be transfected or transformed (in addition to the de-differentiation factors) with at least one additional therapeutic factor. For example, once CiPSCs are isolated, the cells may be transformed with a polynucleotide encoding a therapeutic polypeptide and then implanted or administered to a subject, or may be differentiated to a desired cell type and implanted and delivered to the subject. Under such conditions the polynucleotide is expressed within the subject for delivery of the polypeptide product.

2. Tissue Engineering

CiPSCs and their progeny can be used to make tissue engineered constructions. Tissue engineered constructs may be used for a variety of purposes including as prosthetic devices for the repair or replacement of damaged organs or tissues. They may also serve as in vivo delivery systems for proteins or other molecules secreted by the cells of the construct or as drug delivery systems in general. Tissue engineered constructs also find use as in vitro models of tissue function or as models for testing the effects of various treatments or pharmaceuticals. The biomaterial scaffolds for transplantation of stem cells are described in Willerth, S.M. and Sakiyama-Elbert, S.E., Combining stem cells and biomaterial scaffolds for constructing tissues and cell delivery (July 09, 2008) , StemBook, ed. The Stem Cell Research Community, StemBook, each of which is herein incorporated by reference in its entirety. Tissue engineering technology frequently may involve selection of an appropriate culture substrate to sustain and promote tissue growth. These substrates can be three-dimensional and processable to form scaffolds of a desired shape for the tissue of interest.

Methods for producing tissue engineered constructs and engineered native tissue are described in U.S. Patent No. 6,962,814, which is herein incorporated by reference in its entirety. Methods for using or manufacturing tissue engineered ligaments and tendons are described in U.S. Patent No. 7,914,579, which is herein incorporated by reference in its entirety. U.S. Patent No. 5,716,404 discloses methods and compositions for reconstruction or augmentation of breast tissue using dissociated muscle cells implanted in combination with a polymeric matrix, which is herein incorporated by reference in its entirety. US Patent No. 8,728,495 discloses repair of cartilage using autologous dermal fibroblasts, which is herein incorporated by reference in its entirety. U.S. Published application No. 20090029322 discloses the use of stem cells to form dental tissue for use in making tooth substitute, which is herein incorporated by reference in its entirety. U.S. Published application No. 2006/0019326 discloses cell-seed tissue-engineered polymers for treatment of intracranial aneurysms, which is herein incorporated by reference in its entirety. U.S. Published application No. 2007/0059293 discloses the tissue-engineered constructs (and method for making such constructs) that can be used to replace damaged organs for example kidney, heart, liver, spleen, pancreas, bladder, ureter, and urethra, which is herein incorporated by reference in its entirety.

3. Therapeutic compositions

The CiPSCs can be formulated for administration, delivery or contacting with a subject, tissue, or cell to promote de-differentiation in vivo or in vitro/ex vivo. Additional factors, such as growth factors, other factors that induce differentiation or dedifferentiation, secretion products, immunomodulators, anti-inflammatory agents, regression factors, biologically active compounds that promote innervation, vascularization or enhance the lymphatic network, and drugs, can be incorporated.

The induced pluripotent cells can be administered to a patient by way of a composition that includes a population of CiPSCs or CiPSC progenies alone or on or in a carrier or support structure. In many embodiments, no carrier will be required. The cells can be administered by injection onto or into the site where the cells are required. In these cases, the cells will typically have been washed to remove cell culture media and will be suspended in a physiological buffer. In other embodiments, the cells are provided with or incorporated onto or into a support structure. Support structures may be meshing, solid supports, scaffolds, tubes, porous structures, and/or a hydrogel.

USE OF SMALL REPROGRAMMING FACTORS

The compositions comprising chemical reprogramming factors can be used for tissue regeneration, tissue remodeling and repair, rejuvenation or reversing aging, and inhibiting or reversing fibrosis in vitro and in vivo.

In some cases, chemical reprogramming factors of stage 1 or 2 described herein are formulated for administration, delivery or contacting with a subject, tissue, or cell to promote de-differentiation, regeneration, repair, and rejuvenation in vivo or in vitro/ex vivo. Additional factors, such as growth factors, other factors that induce dedifferentiation or regeneration, secretion products, immunomodulators, anti-inflammatory agents, regression factors, biologically active compounds that promote innervation, vascularization or enhance the lymphatic network, and drugs, can be incorporated.

In one embodiment, the chemical reprogramming factors are administered to a patient by way of a composition that includes all or part of the chemical reprogramming factors for stage 1 or 2 described herein. The chemical reprogramming factor compositions can be administered systemically or by injection onto or into the site where the cells are lost, or tissues are damaged to boost the endogenous repair ability. In some embodiments, no carrier is required. In other embodiments, the compositions can include a pharmaceutically acceptable carrier. The chemical reprogramming factors can also be formulated for sustained release, for example, using microencapsulation. The compositions comprising chemical reprogramming factors can be administered to a patient in a single dose, in multiple doses, in a continuous or intermittent manner to obtain the desired physiological effect, depending on the recipient's physiological conditions. In some embodiments chemical reprogramming factors for stage 1 or 2 described herein are formulated for administration, delivery or contacting with a subject, tissue, or cell to promote rejuvenation. These chemical reprogramming factors can be formulated to prevent the age-associated histological changes and maintain the cells in a younger state in tissues. The rejuvenating effects can be detected by reversion of the epigenetic clock, or metabolic changes, or transcriptomic changes, such as changes in senescence, stress, or inflammation pathways. The compositions comprising chemical reprogramming factors can be administered to a patient in a single dose, in multiple doses, in a continuous or intermittent manner to obtain the desired physiological effect, depending on the recipient's physiological conditions. In some embodiments chemical reprogramming factors for stage 1 or 2 described herein can be formulated for administration, delivery or contacting with a subject, tissue, or cell to inhibit or revise the fibrosis. Fibrosis can be detected by the changes of morphology, epigenome, or metabolic changes, or transcriptomic changes induced by the disease, stress, or inflammations. The compositions comprising chemical reprogramming factors can be administered to a patient in a single dose, in multiple doses, in a continuous or intermittent manner to inhibit or revise the fibrosis, depending on the recipient's physiological conditions

EXAMPLES

The following examples are given for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the present disclosure in any fashion. The present examples, along with the methods described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the disclosure. Changes therein and other uses which are encompassed within the spirit of the disclosure as defined by the scope of the claims will occur to those skilled in the art.

EXAMPLE 1: Methods to convert cells into pluripotent stem cells

This example illustrates methods for converting human somatic cells into pluripotent stem cells according to some embodiments of the present disclosure.

FIG. 1 illustrates a schematic of an exemplary method for converting human somatic cells into pluripotent stem cells. The method has 2 stages. During any of the 3 stages, any cell can proliferate and give rise to progeny cells. In stage 1, a first population of cells comprising somatic cells are contacted with a first composition for about 8-20 days. The first population of cells can comprise somatic cells (e.g., fibroblasts or primary human adult adipose derived mesenchymal stromal cells (hADSCs) ) . During and/or subsequent to contacting the first composition, at least a subset of the cells of the first population of cells or the progenies thereof are converted into a second population of cells. The second population of cells can comprise intermediate plastic state cells. In stage 2, the second population of cells or the progenies thereof are contacted with a second composition for about 6-12 days. During and/or subsequent to contacting the second composition, at least a subset of the cells of the second population of cells or the progenies thereof are converted into a third population of cells. The third population of cells can comprise pluripotent stem cells (e.g., human chemically induced pluripotent stem cells (hCiPSCs) .

In one experiment, when converting hCiPSCs from somatic cell (e.g., hADSCs) , hypoxia with 5%O₂ was used in stage 1 conversion. After stage 1 conversion, cells were incubated with 21%O₂. The induction medium was changed every 3-4 days. hADSCs were seeded at a density of 1 x 10^4 cells per well of a 12-well plate in 15%FBS-DMEM medium and would be changed into stage 1 conversion medium the next day. For stage 1 conversion, Y-27632 (10 μM) , SGC-CBP30 (2 μM) , VTP50469 (0.5 μM) , AKT Kinase Inhibitor (1 μM) , CHIR99021 (2 μM) , E-616452 (10 μM) , TTNPB (2 μM) , SAG (0.5 μM) , EPZ5676 (2 μM) , Ruxolitinib (1 μM) , DZNep (0.2 μM) , JNKIN8 (0.5 μM) , 5-Iodotubercidin (0.5 μM) , CX-4945 (0.5 μM) , Unc0224 (1 μM) , 5-Azacytidine (2 μM) , and BIRB796 (2 μM) were supplemented to the culture medium.

For stage 1 conversion, intermediate plastic state cells induced from hADSCs emerged at day 4-8 and approach 90%confluence at day 8-20. Then the medium was changed into stage 2 conversion medium. For stage 2 conversion, SB590885 (0.5 μM) , VPA (1000 μM) , PD0325901 (1 μM) , IWP2, CHIR99021 (2 μM) , Y-27632 (10 μM) , Tranylcypromine (10 μM) , DZNep (0.2 μM) , and EPZ5676 (2 μM) were supplemented to the culture medium.

EXAMPLE 2: Methods for culturing and analysis of cells

Provided in this example are methods for culturing and analyzing the cells according to some embodiments of the present disclosure.

Cell culture

For culturing Primary human adult adipose derived mesenchymal stromal cells (hADSCs) and human CiPS cells and human ES cells (H1 and H9) can be cultured using the methods described in Nature, 2022, which is herein incorporated by reference in its entirety. Briefly, primary hADSCs can be seeded at a density of 1.5 x 10^6 cells per 100-mm dish and cultured in Mesenchymal Stem Cell Growth Medium 2 under 21%O₂, 5%CO₂ at 37 ℃. The medium can be changed every 2 days and cells can be passaged by 0.25%Trypsin-EDTA before reaching confluence. Primary hADSCs within 4 passages can be used for the induction of CiPS cells. Human CiPS cells and human ES cells (H1 and H9) can be maintained in mTeSR^TM Plus Medium on Matrigel-coated plates (Corning, 354248) under 21%O₂, 5%CO₂ at 37 ℃. The medium can be changed every day and cells can be passaged by ReLeSRTM with split ratios of around 1: 10 to 1: 20 when they reached ～85%confluence. For passaging, detached cell aggregates can be plated in mTeSR^TM Plus Medium supplemented with Y-27632 (10 μM) . After 24 hours, the medium can be replaced with fresh mTeSR^TM Plus Medium without Y-2: 7632.

Isolation and culture of hADSCs

hADSCs can be isolated from adult adipose tissue that is obtained with informed written consent and approval by ethics committee. The procedure can be conducted according to the principles of the Declaration of Helsinki. Briefly: Tissues can be dissociated by collagenase IV and the obtained cells can be plated in a 100-mm dish in Mesenchymal Stem Cell Growth Medium 2 followed by incubation in 21%O₂, 5%CO₂ at 37 ℃. For hCiPSCs induction, hADSCs can be seeded at a density of 1 x 10^4 cells per well of a 12-well plate with 15%FBS-DMEM medium the day before stage 1 induction.

Derivation and culture of human CiPS cell lines

Cells can be dissociated by Accutase (Millipore, SCR005) after 8-12 days’ induction of stage 2 condition, centrifuged at 400 g for 3 min to remove Accutase, and then re-plated at a ratio from 1: 3 to 1: 12 on feeder layers in the modified stage 2 medium: Knockout DMEM supplemented with 1%N2 supplement, 2%B27 supplement, 1%GlutaMAX^TM, 1%NEAA, 1%Penicillin-Streptomycin, 50 μg/ml Vc2p, 2 mg/mL AlbuMAX^TM-II and the small molecules CHIR99021 (1 μM) , PD0325901 (0.5 μM) , IWP-2 (2 μM) , Y-27632 (10 μM) , HRG (20 ng/mL) , and bFGF (100 ng/mL, Origene) .

Immunofluorescence

After fixation with 4%paraformaldehyde at room temperature for 30 min, cells can be blocked by PBS containing 0.1%TritonTM X-100 and 2%donkey serum at 37 ℃ for 1 hour. Primary antibodies incubation with appropriate dilutions can be performed at 4 ℃ overnight in the same buffer. The secondary antibodies can be incubated in PBS containing 2%donkey serum at 4 ℃ overnight. DNA was stained with DAPI solution. For surface marker analysis, PE Anti-Syndecan-1 (CD138) antibodies (Cat#552026; Cat#ab209584) can be used in live-cell immunostaining. The antibodies (1: 200) can be added to the growth medium and then incubated for 2 hours. The cells can be gently washed for three times with prewarmed PBS. Then the cells can be fixed in situ for staining other antibodies.

Flow cytometry and cell sorting

Cells can be dissociated by using 0.25%Trypsin-EDTA and then the cell suspensions can be filtrated through 40 μm cell strainers. FACS analyses can be performed with the BD Cytofix/Cytoperm^TM Fixation/Permeablization Kit (BD Biosciences) according to the manufacturer’s instructions. Flow cytometry can be performed using CytoFlEX S (BeckMan Coulter) . A list of antibodies for the method described herein (e.g., immunofluorescence or flow cytometry/cell sorting) can be found in TABLE 1 below.

TABLE 1: List of antibodies for methods described herein

Teratoma formation

CiPS cells can be harvested by ReLeSR^TM (STEMCELL, Cat#05872) . Approximately 2 x 10^6 cells can be resuspended in Matrigel and then sub-cutaneously injected to the immunodeficient NPG mice. After 6-7 weeks, the teratomas can be obtained and then embedded in paraffin. The paraffin sections can be stained with haematoxylin and eosin.

Reverse transcription-quantitative PCR (qRT-PCR)

Total RNA can be isolated using Direct-zol RNA MiniPrep Kit (Zymo Research, R2053) . cDNA can be synthesized from 0.5-1 μg of total RNA using TransScript First-Strand cDNA Synthesis SuperMix (TransGen Biotech, AT311-03) . qPCR can be performed by using KAPA SYBR FAST qPCR Kit Master Mix (KAPA Biosystems, KM4101) on a CFX ConnectTM Real-Time System (Bio-Rad) . The data can be analyzed using the delta-delta Ct method. GAPDH can be used as a control to normalize the expression of target genes. Primer sequences that can used for qPCR in this study are listed in TABLE 2 below.

TABLE 2. Primers used for qRT-PCR

RNA sequencing

Total RNA can be isolated by using Direct-zol RNA MiniPrep Kit. The NEBNext Ultra RNA Library Prep Kit (NEB England BioLabs, E7775) can be used for RNA sequencing library construction. Fragmented and randomly primed 2 × 150 bp paired-end libraries can be sequenced by using the Illumina HiSeq X Ten system.

Single-cell RNA sequencing (scRNA-seq)

Briefly, cells at different time points can be harvested and resuspended at 1 x 10^6 cells per milliliter in 1 x PBS with 0.04%BSA. Then, the cells can be loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer’s protocol. scRNA-seq libraries can be constructed using the Single Cell 3’ Library and Gel Bead Kit V3.1 (10x Genomics, 1000075) . The libraries can be finally sequenced using the Illumina NovaSeq 6000 sequencer (performed by CapitalBio Technology) .

Whole-genome bisulfite sequencing (WGBS)

Bisulfite conversion of the extracted genome DNA can be performed as previously reported in Guan et al. 2022 (Nature. 2022 May; 605 (7909) : 325-331) , which is herein incorporated by reference in its entirety. The recovered bisulfite-converted DNAs can be constructed into sequencing libraries. For each library, 90 gigabases (Gb) raw data can be obtained by Illumina HiSeq X Ten sequencing system.

EXAMPLE 3: Two-Stage Conversion of Cells

Provided in this example are conversion results following the two-stage reprogramming protocol described in Example 1, according to some embodiments of the present disclosure.

FIG. 2 shows a human chemically induced pluripotent stem cells (hCiPSCs) colony induced by the two-stage protocol, which showed positive staining for expression of OCT4.

While preferred embodiments of the present disclosure have been shown and described herein, it may be obvious to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the disclosure be limited by the specific examples provided within the specification. While the invention has been described with reference to the aforementioned specification, the descriptions and illustrations of the embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions may now occur to those skilled in the art without departing from the disclosure. Furthermore, it shall be understood that all aspects of the disclosure are not limited to the specific depictions, configurations or relative proportions set forth herein which depend upon a variety of conditions and variables. It may be understood that various alternatives to the embodiments described herein may be employed in practicing the present disclosure. It is therefore contemplated that the disclosure shall also cover any such alternatives, modifications, variations, or equivalents. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

A method for producing pluripotent stem cells, comprising:

(a) contacting somatic cells with a first composition comprising one or more of

(1) a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor;

(2) a CBP/p300 bromodomain inhibitor;

(3) a Menin-MLL interaction inhibitor;

(4) a serine-threonine kinase Akt inhibitor;

(5) a glycogen kinase inhibitor;

(6) a TGFβ receptor inhibitor;

(7) a retinoic acid receptor agonist;

(8) an agonist for the G protein-coupled receptor Smoothened;

(9) a Dot1L inhibitor;

(10) a Jak1/Jak2 inhibitor;

(11) an SAH hydrolase inhibitor; or

(12) a c-Jun kinase inhibitor,

thereby obtaining a first cell population; and

(b) contacting the first cell population with a second composition comprising one or more of

(i) a MEK inhibitor;

(ii) a B-Raf inhibitor; or

(iii) a histone deacetylase inhibitor,

thereby obtaining a second cell population comprising pluripotent stem cells.
The method of claim 1, wherein the first composition comprises one or more of

the c-Jun kinase inhibitor,

the CBP/p300 bromodomain inhibitor, or

the serine-threonine kinase Akt inhibitor.
The method of claim 1, wherein the first composition comprises the c-Jun kinase inhibitor, the CBP/p300 bromodomain inhibitor, and the serine-threonine kinase Akt inhibitor.
The method of claim 1, wherein the first composition comprises one or more of:

the ROCK inhibitor;

the CBP/p300 bromodomain inhibitor;

the Menin-MLL interaction inhibitor; or

the serine-threonine kinase Akt inhibitor.
The method of claim 1, wherein the first composition comprises one or more of:

the ROCK inhibitor,

the c-Jun kinase inhibitor, or

the CBP/p300 bromodomain inhibitor.
The method of claim 1, wherein the first composition comprises

(1) the Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor;

(2) the CBP/p300 bromodomain inhibitor;

(3) the Menin-MLL interaction inhibitor;

(4) the serine-threonine kinase Akt inhibitor;

(5) the glycogen kinase inhibitor;

(6) the TGFβ receptor inhibitor;

(7) the retinoic acid receptor agonist;

(8) an agonist for the G protein-coupled receptor Smoothened;

(9) the Dot1L inhibitor;

(10) the Jak1/Jak2 inhibitor;

(11) the SAH hydrolase inhibitor; and

(12) the c-Jun kinase inhibitor.
The method of any one of claims 1-6, wherein the first composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, or a p38 MAPK inhibitor.
The method of any one of claims 1-6, wherein the first composition further comprises a G9a inhibitor or a DNMT inhibitor.
The method of any one of claims 1-6, wherein the first composition further comprises an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or both.
The method of any one of claims 1-9, wherein the second composition further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, a ROCK inhibitor, an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor.
The method of any one of claims 1-9, wherein the second composition further comprises one or more of an inhibitor of histone demethylation, a Dot1L inhibitor, or an SAH hydrolase inhibitor.
The method of any one of claims 1-9, wherein the second composition further comprises one or more of a Wnt inhibitor, a glycogen kinase inhibitor, or a ROCK inhibitor.
The method of any one of claims 1-12, wherein the first cell population comprises intermediate plastic state cells that express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.
The method of claim 13, wherein the intermediate plastic state cells express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1.
The method of claim 13 or 14, wherein the intermediate plastic state cells further express one or more of FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.
The method of any one of claims 1-15, wherein the pluripotent stem cells express one or more of OCT4, SOX2, or NANOG.
The method of any one of claims 1-16, wherein the pluripotent stem cells further express one or more of FGF4, ZFP57, DPPA5, REX1, DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DPPA5, DNMT3L, REX1, or UTF1.
The method of any one of claims 1-16, wherein the pluripotent stem cells further express one or more of FGF4, ZFP57, DPPA5, or REX1.
The method of any one of claims 1-18, wherein the pluripotent stem cells further express one or more of DPPA4, TDGF1, TRA-1-60, TRA-1-81, SSEA4, KLF4, KLF17, DPPA3, DPPA5, DNMT3L, REX1, or UTF1.
The method of any one of claims 1-19, wherein the somatic cells comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) .
The method of any one of claims 1-19, wherein the somatic cells comprise fibroblasts.
The method of any one of claims 1-21, wherein the method converts the somatic cells into the pluripotent stem cells within less about 50 days.
The method of any one of claims 1-21, wherein the method converts the somatic cells into the pluripotent stem cells within at most about 32 days.
The method of any one of claims 1-21, wherein the method converts the somatic cells into the pluripotent stem cells within at most about 24 days.
The method of any one of claims 1-21, wherein the method converts the somatic cells into the pluripotent stem cells within at most about 14 days.
The method of any one of claims 1-25, wherein the somatic cells comprise a genetic modification.
The method of claim 26, wherein the pluripotent stem cells comprise the genetic modification.
The method of claim 26 or 27, wherein the genetic modification comprises an exogenous nucleic acid sequence.
The method of claim 28, wherein the exogenous nucleic acid sequence encodes a polypeptide.
The method of claim 28, wherein the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule.
The method of claim 26 or 27, wherein the genetic modification comprises alteration of a genomic sequence.
The method of any one of claims 26-31, wherein the genetic modification reduces immunogenicity of the pluripotent stem cells.
The method of any one of claims 1-32, wherein the ROCK inhibitor comprises Y-27632 or thiazovivin.
The method of claim 33, wherein the ROCK inhibitor comprises Y-27632.
The method of claim 34, wherein Y-27632 is present at about 1 μM to about 100 μM within the first composition or the second composition.
The method of claim 34, wherein Y-27632 is present at about 2 μM to about 50 μM within the first composition or the second composition.
The method of claim 34, wherein Y-27632 is present at about 4 μM to about 25 μM within the first composition or the second composition.
The method of claim 34, wherein Y-27632 is present at about 10 μM within the first composition or the second composition.
The method of any one of claims 1-38, wherein the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409.
The method of any one of claims 1-38, wherein the CBP/p300 bromodomain inhibitor comprises SGC-CBP30.
The method of claim 40, wherein SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM within the first composition.
The method of claim 40, wherein SGC-CBP30 is present at about 0.4 μM to about 10 μM within the first composition.
The method of claim 40, wherein SGC-CBP30 is present at about 0.8 μM to about 5 μM within the first composition.
The method of claim 40, wherein SGC-CBP30 is present at about 2 μM within the first composition.
The method of any one of claims 1-44, wherein the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4.
The method of any one of claims 1-44, wherein the Menin-MLL interaction inhibitor comprises VTP50469.
The method of claim 46, wherein VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM within the first composition.
The method of claim 46, wherein VTP50469 is present at about 0.1 μM to about 2.5 μM within the first composition.
The method of claim 46, wherein VTP50469 is present at about 0.2 μM to about 1.25 μM within the first composition.
The method of claim 46, wherein VTP50469 is present at about 0.5 μM within the first composition.
The method of any one of claims 1-50, wherein the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor.
The method of claim 51, wherein AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM within the first composition.
The method of claim 51, wherein AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM within the first composition.
The method of claim 51, wherein AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM within the first composition.
The method of claim 51, wherein AKT Kinase Inhibitor is present at about 1 μM within the first composition.
The method of any one of claims 1-55, wherein the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014.
The method of any one of claims 1-55, wherein the glycogen kinase inhibitor comprises CHIR99021.
The method of claim 57, wherein CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM within the first composition or the second composition.
The method of claim 57, wherein CHIR99021 is present at about 1 μM to about 25 μM within the first composition or the second composition.
The method of claim 57, wherein CHIR99021 is present at about 2 μM to about 12.5 μM within the first composition.
The method of claim 57, wherein CHIR99021 is present at about 5 μM within the first composition.
The method of claim 57, wherein CHIR99021 is present at about 0.75 μM to about 5 μM within the second composition.
The method of claim 57, wherein CHIR99021 is present at about 1 μM to about 3 μM within the second composition.
The method of claim 57, wherein CHIR99021 is present at about 2 μM within the second composition.
The method of any one of claims 1-64, wherein the TGFβ receptor inhibitor is an ALK5 inhibitor.
The method of any one of claims 1-64, wherein the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334.
The method of any one of claims 1-64, wherein the TGFβ receptor inhibitor comprises E-616452.
The method of claim 67, wherein E-616452 is present at about 1 micromolar (μM) to about 100 μM within the first composition.
The method of claim 67, wherein E-616452 is present at about 2 μM to about 50 μM within the first composition.
The method of claim 67, wherein E-616452 is present at about 4 μM to about 25 μM within the first composition.
The method of claim 67, wherein E-616452 is present at about 10 μM within the first composition.
The method of any one of claims 1-71, wherein the RAR agonist comprises TTNPB, Ch55, or AM580.
The method of any one of claims 1-71, wherein the RAR agonist comprises TTNPB.
The method of claim 73, wherein TTNPB is present at about 0.2 μM to about 20 μM within the first composition.
The method of claim 73, wherein TTNPB is present at about 0.4 μM to about 10 μM within the first composition.
The method of claim 73, wherein TTNPB is present at about 0.8 μM to about 5 μM within the first composition.
The method of claim 73, wherein TTNPB is present at about 2 μM within the first composition.
The method of any one of claims 1-77, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH.
The method of any one of claims 1-77, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG.
The method of claim 79, wherein SAG is present at about 0.05 micromolar (μM) to about 5 μM within the first composition.
The method of claim 79, wherein SAG is present at about 0.1 μM to about 2.5 μM within the first composition.
The method of claim 79, wherein SAG is present at about 0.2 μM to about 1.25 μM within the first composition.
The method of claim 79, wherein SAG is present at about 0.5 μM within the first composition.
The method of any one of claims 1-83, wherein the Dot1L inhibitor comprises EPZ004777 or EPZ5676.
The method of any one of claims 1-83, wherein the Dot1L inhibitor comprises the EPZ5676.
The method of claim 85, wherein EPZ5676 is present at about 0.2 μM to about 20 μM within the first composition or the second composition.
The method of claim 85, wherein EPZ5676 is present at about 0.4 μM to about 10 μM within the first composition or the second composition.
The method of claim 85, wherein EPZ5676 is present at about 0.8 μM to about 5 μM within the first composition or the second composition.
The method of claim 85, wherein EPZ5676 is present at about 2 μM within the first composition or the second composition.
The method of any one of claims 1-89, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib.
The method of any one of claims 1-89, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib.
The method of claim 91, wherein Ruxolitinib is present at about 0.1 μM to about 10 μM within the first composition.
The method of claim 91, wherein Ruxolitinib is present at about 0.2 μM to about 5 μM within the first composition.
The method of claim 91, wherein Ruxolitinib is present at about 0.4 μM to about 2.5 μM within the first composition.
The method of claim 91, wherein Ruxolitinib is present at about 1 μM within the first composition.
The method of any one of claims 1-95, wherein the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA.
The method of any one of claims 1-95, wherein the SAH hydrolase inhibitor comprises DZNep.
The method of claim 97, wherein DZNep is present at about 0.02 μM to about 2 μM within the first composition or the second composition.
The method of claim 97, wherein DZNep is present at about 0.04 μM to about 1 μM within the first composition or the second composition.
The method of claim 97, wherein DZNep is present at about 0.08 μM to about 0.5 μM within the first composition or the second composition.
The method of claim 97, wherein DZNep is present at about 0.2 μM with the first composition or the second composition.
The method of any one of claims 1-101, wherein the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12.
The method of any one of claims 1-101, wherein the c-Jun kinase inhibitor comprises JNKIN8.
The method of claim 103, wherein JNKIN8 is present at about 0.05 micromolar (μM) to about 5 μM within the first composition.
The method of claim 103, wherein JNKIN8 is present at about 0.1 μM to about 2.5 μM within the first composition.
The method of claim 103, wherein JNKIN8 is present at about 0.2 μM to about 1.25 μM within the first composition.
The method of claim 103, wherein JNKIN8 is present at about 0.5 μM within the first composition.
The method of any one of claims 1-107, wherein the MEK inhibitor comprises PD0325901, AZD8330, or TAK-733.
The method of any one of claims 1-107, wherein the MEK inhibitor comprises PD0325901.
The method of claim 109, wherein PD0325901 is present at about 0.1 μM to about 10 μM with the second composition.
The method of claim 109, wherein PD0325901 is present at about 0.2 μM to about 5 μM with the second composition.
The method of claim 109, wherein PD0325901 is present at about 0.4 μM to about 2.5 μM with the second composition.
The method of claim 109, wherein PD0325901 is present at about 1 μM with the second composition.
The method of any one of claims 1-113, wherein the B-Raf inhibitor comprises SB590885, Vemurafenib, RAF265, or PLX4720.
The method of any one of claims 1-113, wherein the B-Raf inhibitor comprises SB590885.
The method of claim 115, wherein SB590885 is present at about 0.05 μM to about 5 μM with the second composition.
The method of claim 115, wherein SB590885 is present at about 0.1 μM to about 2.5μM with the second composition.
The method of claim 115, wherein SB590885 is present at about 0.2 μM to about 1.25 μM with the second composition.
The method of claim 115, wherein SB590885 is present at about 0.5 μM with the second composition.
The method of any one of claims 1-119, wherein the histone deacetylase inhibitor comprises valproic acid (VPA) , LMK235, MS275, or HDACi IV.
The method of any one of claims 1-119, wherein the histone deacetylase inhibitor comprises valproic acid (VPA) .
The method of claim 121, wherein VPA is present at about 0.1 millimolar (mM) to about 10 mM within the second composition.
The method of claim 121, wherein VPA is present at about 0.2 mM to about 5 mM within the second composition.
The method of claim 121, wherein VPA is present at about 0.4 mM to about 2.5 mM within the second composition.
The method of claim 121, wherein VPA is present at about 1 mM within the second composition.
The method of any one of claims 1-125, wherein the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702.
The method of any one of claims 1-125, wherein the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) .
The method of claim 127, wherein 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM within the first composition.
The method of claim 127, wherein 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM within the first composition.
The method of claim 127, wherein 5-ITU is present at about 0.2 micromolar μM to about 1 μM within the first composition.
The method of claim 127, wherein the 5-ITU is present at about 0.5 μM within the first composition.
The method of any one of claims 1-131, wherein the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid.
The method of any one of claims 1-131, wherein the casein kinase 2 inhibitor comprises the CX-4945.
The method of claim 133, wherein CX-4945 is present at about 0.1 μM about 10 μM within the first composition.
The method of claim 133, wherein CX-4945 is present at about 0.2 μM about 5 μM within the first composition.
The method of claim 133, wherein CX-4945 is present at about 0.5 μM about 1 μM within the first composition.
The method of any one of claims 1-136, wherein the G9a inhibitor comprises Unc0224, Unc0638, Unc0642, or Bix01294.
The method of any one of claims 1-136, wherein the G9a inhibitor comprises Unc0224.
The method of claim 138, wherein Unc0224 is present at about 0.1 μM to about 10 μM within the first composition.
The method of claim 138, wherein Unc0224 is present at about 0.2 μM to about 5 μM within the first composition.
The method of claim 138, wherein Unc0224 is present at about 0.5 μM to about 2 μM within the first composition.
The method of claim 138, wherein Unc0224 is present at about 1 μM within the first composition.
The method of any one of claims 1-142, wherein the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027.
The method of any one of claims 1-142, wherein the DNMT inhibitor comprises 5-Azacytidine.
The method of claim 144, wherein 5-Azacytidine is present at about 0.2 μM to about 20 μM within the first composition.
The method of claim 144, wherein 5-Azacytidine is present at about 0.5 μM to about 10 μM within the first composition.
The method of claim 144, wherein 5-Azacytidine is present at about 1 μM to about 5 μM within the first composition.
The method of claim 144, wherein 5-Azacytidine is present at about 2 μM within the first composition.
The method of any one of claims 1-148, wherein the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190.
The method of any one of claims 1-148, wherein the p38 MAPK inhibitor comprises BIRB796.
The method of claim 150, wherein BIRB796 is present at about 0.2 μM to about 20 μM within the first composition.
The method of claim 150, wherein BIRB796 is present at about 0.4 μM to about 10 μM within the first composition.
The method of claim 150, wherein BIRB796 is present at about 0.8 μM to about 5 μM within the first composition.
The method of claim 150, wherein BIRB796 is present at about 2 μM within the first composition.
The method of any one of claims 10-154, wherein the inhibitor of histone demethylation comprises Tranylcypromine.
The method of claim 155, wherein Tranylcypromine is present at about 1 μM to about 100 μM within the second composition.
The method of claim 155, wherein Tranylcypromine is present at about 2 μM to about 50 μM within the second composition.
The method of claim 155, wherein Tranylcypromine is present at about 4 μM to about 25 μM within the second composition.
The method of claim 155, wherein Tranylcypromine is present at about 10 μM within the second composition.
The method of any one of claims 10-159, wherein the Wnt inhibitor comprises IWR-1 or IWP-2.
The method of claim 160, wherein the Wnt inhibitor comprises IWR-1.
The method of claim 160, wherein the Wnt inhibitor comprises IWP-2.
The method of claim 162, wherein IWP-2 is present at about 0.2 μM to about 20 μM within the second composition.
The method of claim 162, wherein IWP-2 is present at about 0.4 μM to about 10 μM within the second composition.
The method of claim 162, wherein IWP-2 is present at about 0.8 μM to about 5 μM within the second composition.
The method of claim 162, wherein IWP-2 is present at about 2 μM within the second composition.
A method for reprogramming somatic cells, comprising contacting the somatic cells with a composition comprising a c-Jun kinase inhibitor or a CBP/p300 bromodomain inhibitor.
The method of claim 167, wherein the composition further comprises a serine-threonine kinase Akt inhibitor.
A method for reprogramming somatic cells, comprising contacting the somatic cells with a composition comprising one or more of a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor.
The method of any one of claims 167-169, wherein the composition further comprises a ROCK inhibitor or a Menin-MLL interaction inhibitor.
The method of any one of claims 167-170, wherein the composition further comprises one or more of:

(1) a glycogen kinase inhibitor;

(2) a TGFβ receptor inhibitor;

(3) a retinoic acid receptor agonist;

(4) an agonist for the G protein-coupled receptor Smoothened;

(5) a Dot1L inhibitor;

(6) a Jak1/Jak2 inhibitor; or

(7) an SAH hydrolase inhibitor.
The method of any one of claims 167-171, wherein the composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, a p38 MAPK inhibitor.
The method of any one of claims 167-171, wherein the composition further comprises a G9a inhibitor or a DNMT inhibitor.
The method of any one of claims 167-171, wherein the composition comprises the adenosine kinase inhibitor, the casein kinase 2 inhibitor, or both.
The method of any one of claims 167-171, wherein the method induces conversion of the somatic cells into intermediate plastic state cells that express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, LEF1, FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.
The method of claim 175, wherein the intermediate plastic state cells express one or more of MSX1, HOXB9, WT1, GATA2, HMGA2, or LEF1.
The method of claim 176, wherein the intermediate plastic state cells further express one or more of FGF9, HOXA9, HOZXA1, PTCH1, HOXA5, CCND2, SDC1, TBX3, BMP4, or IGF2.
The method of any one of claims 167-177, wherein the somatic cells comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) .
The method of any one of claims 167-177, wherein the somatic cells comprise fibroblasts.
The method of any one of claims 167-179, wherein the somatic cells comprise a genetic modification.
The method of claim 180, wherein the intermediate plastic state cells comprise the genetic modification.
The method of claim 180 or 181, wherein the genetic modification comprises an exogenous nucleic acid sequence.
The method of claim 182, wherein the exogenous nucleic acid sequence encodes a polypeptide.
The method of claim 182, wherein the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule.
The method of any one of claims 180-184, wherein the genetic modification comprises alteration of a genomic sequence.
The method of any one of claims 180-185, wherein the genetic modification reduces immunogenicity of the intermediate plastic state cells.
The method of any one of claims 167-186, wherein the ROCK inhibitor comprises Y-27632 or thiazovivin.
The method of any one of claims 167-186, wherein the ROCK inhibitor comprises Y-27632.
The method of claim 188, wherein Y-27632 is present at about 1 μM to about 100 μM within the composition.
The method of claim 188, wherein Y-27632 is present at about 2 μM to about 50 μM within the composition.
The method of claim 188, wherein Y-27632 is present at about 4 μM to about 25 μM within the composition.
The method of claim 188, wherein Y-27632 is present at about 10 μM within the composition.
The method of any one of claims 167-192, wherein the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409.
The method of any one of claims 167-192, wherein the CBP/p300 bromodomain inhibitor comprises SGC-CBP30.
The method of claim 194, wherein SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM within the composition.
The method of claim 194, wherein SGC-CBP30 is present at about 0.4 μM to about 10 μM within the composition.
The method of claim 194, wherein SGC-CBP30 is present at about 0.8 μM to about 5 μM within the composition.
The method of claim 194, wherein SGC-CBP30 is present at about 2 μM within the composition.
The method of any one of claims 167-198, wherein the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4.
The method of any one of claims 167-198, wherein the Menin-MLL interaction inhibitor comprises VTP50469.
The method of claim 200, wherein VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM within the composition.
The method of claim 200, wherein VTP50469 is present at about 0.1 μM to about 2.5 μM within the composition.
The method of claim 200, wherein VTP50469 is present at about 0.2 μM to about 1.25 μM within the composition.
The method of claim 200, wherein VTP50469 is present at about 0.5 μM within the composition.
The method of any one of claims 167-198, wherein the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor.
The method of claim 205, wherein AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM within the composition.
The method of claim 205, wherein AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM within the composition.
The method of claim 205, wherein AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM within the composition.
The method of claim 205, wherein AKT Kinase Inhibitor is present at about 1 μM within the composition.
The method of any one of claims 167-209, wherein the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014.
The method of any one of claims 167-209, wherein the glycogen kinase inhibitor comprises CHIR99021.
The method of claim 211, wherein CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM within the composition.
The method of claim 211, wherein CHIR99021 is present at about 1 μM to about 25 μM within the composition.
The method of claim 211, wherein CHIR99021 is present at about 2 μM to about 12.5 μM within the composition.
The method of claim 211, wherein CHIR99021 is present at about 5 μM within the composition.
The method of any one of claims 167-215, wherein the TGFβ receptor inhibitor is an ALK5 inhibitor.
The method of any one of claims 167-215, wherein the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334.
The method of any one of claims 167-215, wherein the TGFβ receptor inhibitor comprises E-616452.
The method of claim 218, wherein E-616452 is present at about 1 micromolar (μM) to about 100 μM within the composition.
The method of claim 218, wherein E-616452 is present at about 2 μM to about 50 μM within the composition.
The method of claim 218, wherein E-616452 is present at about 4 μM to about 25 μM within the composition.
The method of claim 218, wherein E-616452 is present at about 10 μM within the composition.
The method of any one of claims 167-222, wherein the RAR agonist comprises TTNPB, Ch55, or AM580.
The method of any one of claims 167-222, wherein the RAR agonist comprises TTNPB.
The method of claim 224, wherein TTNPB is present at about 0.2 μM to about 20 μM within the composition.
The method of claim 224, wherein TTNPB is present at about 0.4 μM to about 10 μM within the composition.
The method of claim 224, wherein TTNPB is present at about 0.8 μM to about 5 μM within the composition.
The method of claim 224, wherein TTNPB is present at about 2 μM within the composition.
The method of any one of claims 167-228, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH.
The method of any one of claims 167-228, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG.
The method of claim 230, wherein SAG is present at about 0.05 micromolar (μM) to about 5 μM within the composition.
The method of claim 230, wherein SAG is present at about 0.1 μM to about 2.5 μM within the composition.
The method of claim 230, wherein SAG is present at about 0.2 μM to about 1.25 μM within the composition.
The method of claim 230, wherein SAG is present at about 0.5 μM within the composition.
The method of any one of claims 167-234, wherein the Dot1L inhibitor comprises EPZ004777 or EPZ5676.
The method of any one of claims 167-234, wherein the Dot1L inhibitor comprises the EPZ5676.
The method of claim 236, wherein EPZ5676 is present at about 0.2 μM to about 20 μM within the composition.
The method of claim 236, wherein EPZ5676 is present at about 0.4 μM to about 10 μM within the composition.
The method of claim 236, wherein EPZ5676 is present at about 0.8 μM to about 5 μM within the composition.
The method of claim 236, wherein EPZ5676 is present at about 2 μM within the composition.
The method of any one of claims 167-240, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib.
The method of any one of claims 167-240, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib.
The method of claim 242, wherein Ruxolitinib is present at about 0.1 μM to about 10 μM within the composition.
The method of claim 242, wherein Ruxolitinib is present at about 0.2 μM to about 5 μM within the composition.
The method of claim 242, wherein Ruxolitinib is present at about 0.4 μM to about 2.5 μM within the composition.
The method of claim 242, wherein Ruxolitinib is present at about 1 μM within the composition.
The method of any one of claims 167-246, wherein the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA.
The method of any one of claims 167-246, wherein the SAH hydrolase inhibitor comprises DZNep.
The method of claim 248, wherein DZNep is present at about 0.02 μM to about 2 μM within the composition.
The method of claim 248, wherein DZNep is present at about 0.04 μM to about 1 μM within the composition.
The method of claim 248, wherein DZNep is present at about 0.08 μM to about 0.5 μM within the composition.
The method of claim 248, wherein DZNep is present at about 0.2 μM with the composition.
The method of any one of claims 167-252, wherein the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12.
The method of any one of claims 167-252, wherein the c-Jun kinase inhibitor comprises JNKIN8.
The method of claim 254, wherein JNKIN is present at about 0.05 micromolar (μM) to about 5 μM within the composition.
The method of claim 254, wherein JNKIN is present at about 0.1 μM to about 2.5 μM within the composition.
The method of claim 254, wherein JNKIN is present at about 0.2 μM to about 1.25 μM within the composition.
The method of claim 254, wherein JNKIN is present at about 0.5 μM within the composition.
The method of any one of claims 167-258, wherein the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702.
The method of any one of claims 167-258, wherein the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) .
The method of claim 260, wherein 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM within the composition.
The method of claim 260, wherein 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM within the composition.
The method of claim 260, wherein 5-ITU is present at about 0.2 micromolar μM to about 1 μM within the composition.
The method of claim 260, wherein the 5-ITU is present at about 0.5 μM within the composition.
The method of any one of claims 167-264, wherein the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid.
The method of any one of claims 167-264, wherein the casein kinase 2 inhibitor comprises the CX-4945.
The method of claim 266, wherein CX-4945 is present at about 0.1 μM about 10 μM within the composition.
The method of claim 266, wherein CX-4945 is present at about 0.2 μM about 5 μM within the composition.
The method of claim 266, wherein CX-4945 is present at about 0.5 μM about 1 μM within the composition.
The method of any one of claims 167-269, wherein the G9a inhibitor comprises Unc0224, Unc0638, Unc0642, or Bix01294.
The method of any one of claims 167-269, wherein the G9a inhibitor comprises Unc0224.
The method of claim 271, wherein Unc0224 is present at about 0.1 μM to about 10 μM within the composition.
The method of claim 271, wherein Unc0224 is present at about 0.2 μM to about 5 μM within the composition.
The method of claim 271, wherein Unc0224 is present at about 0.5 μM to about 2 μM within the composition.
The method of claim 271, wherein Unc0224 is present at about 1 μM within the composition.
The method of any one of claims 167-275, wherein the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027.
The method of any one of claims 167-275, wherein the DNMT inhibitor comprises 5-Azacytidine.
The method of claim 277, wherein 5-Azacytidine is present at about 0.2 μM to about 20 μM within the composition.
The method of claim 277, wherein 5-Azacytidine is present at about 0.5 μM to about 10 μM within the composition.
The method of claim 277, wherein 5-Azacytidine is present at about 1 μM to about 5 μM within the composition.
The method of claim 277, wherein 5-Azacytidine is present at about 2 μM within the composition.
The method of any one of claims 167-281, wherein the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190.
The method of any one of claims 167-281, wherein the p38 MAPK inhibitor comprises BIRB796.
The method of claim 283, wherein BIRB796 is present at about 0.2 μM to about 20 μM within the composition.
The method of claim 283, wherein BIRB796 is present at about 0.4 μM to about 10 μM within the composition.
The method of claim 283, wherein BIRB796 is present at about 0.8 μM to about 5 μM within the composition.
The method of claim 283, wherein BIRB796 is present at about 2 μM within the composition.
A composition, comprising:

(1) one or more of a G9a inhibitor or a DNMT inhibitor; and

(2) one or more of a glycogen kinase inhibitor, a TGFβ receptor inhibitor, a retinoic acid receptor agonist, a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a CBP/p300 bromodomain inhibitor, a Menin-MLL interaction inhibitor, a serine-threonine kinase Akt inhibitor, an agonist for the G protein-coupled receptor Smoothened, a Dot1L inhibitor, an SAH hydrolase inhibitor, a c-Jun kinase inhibitor, an adenosine kinase inhibitor, a casein kinase 2 inhibitor, or a p38 MAPK inhibitor.
The composition of claim 288, comprising: the glycogen kinase inhibitor, the TGFβ receptor inhibitor, the retinoic acid receptor agonist, the ROCK inhibitor, the CBP/p300 bromodomain inhibitor, the Menin-MLL interaction inhibitor, the serine-threonine kinase Akt inhibitor, the agonist for the G protein-coupled receptor Smoothened, and the Dot1L inhibitor.
The composition of claim 288 or 289, comprising the SAH hydrolase inhibitor, and the c-Jun kinase inhibitor.
The composition of any one of claims 288-290, comprising the G9a inhibitor, the DNMT inhibitor, and the p38 MAPK inhibitor.
The composition of any one of claims 288-290, wherein the ROCK inhibitor comprises Y-27632 or thiazovivin.
The composition of any one of claims 288-290, wherein the ROCK inhibitor comprises Y-27632.
The composition of claim 293, wherein Y-27632 is present at about 1 μM to about 100 μM in the composition.
The composition of claim 293, wherein Y-27632 is present at about 2 μM to about 50 μM in the composition.
The composition of claim 293, wherein Y-27632 is present at about 4 μM to about 25 μM in the composition.
The composition of claim 293, wherein Y-27632 is present at about 10 μM in the composition.
The composition of any one of claims 288-297, wherein the CBP/p300 bromodomain inhibitors comprises SGC-CBP30, I-CBP112, GNE272, or GNE409.
The composition of any one of claims 288-297, wherein the CBP/p300 bromodomain inhibitor comprises SGC-CBP30.
The composition of claim 299, wherein SGC-CBP30 is present at about 0.2 micromolar (μM) to about 20 μM in the composition wherein SGC-CBP30 is present at about 0.4 μM to about 10 μM in the composition.
The composition of claim 299, wherein SGC-CBP30 is present at about 0.8 μM to about 5 μM in the composition.
The composition of claim 299, wherein SGC-CBP30 is present at about 2 μM in the composition.
The composition of any one of claims 288-302, wherein the Menin-MLL interaction inhibitor comprises VTP50469, MI3454, or WDR5-IN-4.
The composition of any one of claims 288-302, wherein the Menin-MLL interaction inhibitor comprises VTP50469.
The composition of claim 304, wherein VTP50469 is present at about 0.05 micromolar (μM) to about 5 μM in the composition.
The composition of claim 304, wherein VTP50469 is present at about 0.1 μM to about 2.5 μM in the composition.
The composition of claim 304, wherein VTP50469 is present at about 0.2 μM to about 1.25 μM in the composition.
The composition of claim 304, wherein VTP50469 is present at about 0.5 μM in the composition.
The composition of any one of claims 288-308, wherein the serine-threonine kinase Akt inhibitor comprises AKT Kinase Inhibitor.
The composition of claim 309, wherein AKT Kinase Inhibitor is present at about 0.1 μM to about 10 μM in the composition.
The composition of claim 309, wherein AKT Kinase Inhibitor is present at about 0.2 μM to about 5 μM in the composition.
The composition of claim 309, wherein AKT Kinase Inhibitor is present at about 0.5 μM to about 2 μM in the composition.
The composition of claim 309, wherein AKT Kinase Inhibitor is present at about 1 μM in the composition.
The composition of any one of claims 288-313, wherein the glycogen kinase inhibitor comprises CHIR99021 or CHIR98014.
The composition of any one of claims 288-313, wherein the glycogen kinase inhibitor comprises CHIR99021.
The composition of claim 315, wherein CHIR99021 is present at about 0.5 micromolar (μM) to about 50 μM in the composition.
The composition of claim 315, wherein CHIR99021 is present at about 1 μM to about 25 μM in the composition.
The composition of claim 315, wherein CHIR99021 is present at about 2 μM to about 12.5 μM in the composition.
The composition of claim 315, wherein CHIR99021 is present at about 5 μM in the composition.
The composition of any one of claims 288-319, wherein the TGFβ receptor inhibitor is an ALK5 inhibitor.
The composition of any one of claims 288-319, wherein the TGFβ receptor inhibitor comprises E-616452, A 83-01, SB431542, SB 505124, GW 788388, dorsomorphin, or SB 525334.
The composition of any one of claims 288-319, wherein the TGFβ receptor inhibitor comprises E-616452.
The composition of claim 322, wherein E-616452 is present at about 1 micromolar (μM) to about 100 μM in the composition.
The composition of claim 322, wherein E-616452 is present at about 2 μM to about 50 μM in the composition.
The composition of claim 322, wherein E-616452 is present at about 4 μM to about 25 μM in the composition.
The composition of claim 322, wherein E-616452 is present at about 10 μM in the composition.
The composition of any one of claims 288-326, wherein the RAR agonist comprises TTNPB, Ch55, or AM580.
The composition of any one of claims 288-326, wherein the RAR agonist comprises TTNPB.
The composition of claim 328, wherein TTNPB is present at about 0.2 μM to about 20 μM in the composition.
The composition of claim 328, wherein TTNPB is present at about 0.4 μM to about 10 μM in the composition.
The composition of claim 328, wherein TTNPB is present at about 0.8 μM to about 5 μM in the composition.
The composition of claim 328, wherein TTNPB is present at about 2 μM in the composition.
The composition of any one of claims 288-332, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG, Purmorphamine, Hh-Ag1.5, or human SHH.
The composition of any one of claims 288-332, wherein the agonist for the G protein-coupled receptor Smoothened comprises SAG.
The composition of claim 334, wherein SAG is present at about 0.05 micromolar (μM) to about 5 μM in the composition.
The composition of claim 334, wherein SAG is present at about 0.1 μM to about 2.5 μM in the composition.
The composition of claim 334, wherein SAG is present at about 0.2 μM to about 1.25 μM in the composition.
The composition of claim 334, wherein SAG is present at about 0.5 μM in the composition.
The composition of any one of claims 288-338, wherein the Dot1L inhibitor comprises EPZ004777 or EPZ5676.
The composition of any one of claims 288-338, wherein the Dot1L inhibitor comprises the EPZ5676.
The composition of claim 340, wherein EPZ5676 is present at about 0.2 μM to about 20 μM in the composition.
The composition of claim 340, wherein EPZ5676 is present at about 0.4 μM to about 10 μM in the composition.
The composition of claim 340, wherein EPZ5676 is present at about 0.8 μM to about 5 μM in the composition.
The composition of claim 340, wherein EPZ5676 is present at about 2 μM in the composition.
The composition of any one of claims 288-344, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib, Tofacitinib, AZD1480, Baricitinib, or Fedratinib.
The composition of any one of claims 288-344, wherein the Jak1/Jak2 inhibitor comprises Ruxolitinib.
The composition of claim 346, wherein Ruxolitinib is present at about 0.1 μM to about 10 μM in the composition.
The composition of claim 346, wherein Ruxolitinib is present at about 0.2 μM to about 5 μM in the composition.
The composition of claim 346, wherein Ruxolitinib is present at about 0.4 μM to about 2.5 μM in the composition.
The composition of claim 346, wherein Ruxolitinib is present at about 1 μM in the composition.
The composition of any one of claims 288-350, wherein the SAH hydrolase inhibitor comprises DZNep, NepA, Adox, or DZA.
The composition of any one of claims 288-350, wherein the SAH hydrolase inhibitor comprises DZNep.
The composition of claim 352, wherein DZNep is present at about 0.02 μM to about 2 μM in the composition.
The composition of claim 352, wherein DZNep is present at about 0.04 μM to about 1 μM in the composition.
The composition of claim 352, wherein DZNep is present at about 0.08 μM to about 0.5 μM in the composition.
The composition of claim 352, wherein DZNep is present at about 0.2 μM in the composition.
The composition of any one of claims 288-356, wherein the c-Jun kinase inhibitor comprises JNKIN8, JNKIN7, JNKIN5, or JNKIN12.
The composition of any one of claims 288-356, wherein the c-Jun kinase inhibitor comprises JNKIN8.
The composition of claim 358, wherein JNKIN is present at about 0.05 micromolar (μM) to about 5 μM in the composition.
The composition of claim 358, wherein JNKIN is present at about 0.1 μM to about 2.5 μM in the composition.
The composition of claim 358, wherein JNKIN is present at about 0.2 μM to about 1.25 μM in the composition.
The composition of claim 358, wherein JNKIN is present at about 0.5 μM in the composition.
The composition of any one of claims 288-362, wherein the adenosine kinase inhibitor comprises 5-Iodotubercidin or ABT 702.
The composition of any one of claims 288-362, wherein the adenosine kinase inhibitor 5-Iodotubercidin (5-ITU) .
The composition of claim 364, wherein 5-ITU is present at about 0.05 micromolar (μM) to about 5 μM in the composition.
The composition of claim 364, wherein 5-ITU is present at about 0.1 micromolar μM to about 2.5 μM in the composition.
The composition of claim 364, wherein 5-ITU is present at about 0.2 micromolar μM to about 1 μM in the composition.
The composition of claim 364, wherein the 5-ITU is present at about 0.5 μM in the composition.
The composition of any one of claims 288-368, wherein the casein kinase 2 inhibitor comprises CX-4945, TPP 22, or Ellagic acid.
The composition of any one of claims 288-368, wherein the casein kinase 2 inhibitor comprises the CX-4945.
The composition of claim 370, wherein CX-4945 is present at about 0.1 μM about 10 μM in the composition.
The composition of claim 370, wherein CX-4945 is present at about 0.2 μM about 5 μM in the composition.
The composition of claim 370, wherein CX-4945 is present at about 0.5 μM about 1 μM in the composition.
The composition of any one of claims 288-373, wherein the G9a inhibitor comprises Unc0224, Unc0638, Unc0642, or Bix01294.
The composition of any one of claims 288-373, wherein the G9a inhibitor comprises Unc0224.
The composition of claim 375, wherein Unc0224 is present at about 0.1 μM to about 10 μM in the composition.
The composition of claim 375, wherein Unc0224 is present at about 0.2 μM to about 5 μM in the composition.
The composition of claim 375, wherein Unc0224 is present at about 0.5 μM to about 2 μM in the composition.
The composition of claim 375, wherein Unc0224 is present at about 1 μM in the composition.
The composition of any one of claims 288-379, wherein the DNMT inhibitor comprises 5-Azacytidine, Decitabine, RG108, or SGI-1027.
The composition of any one of claims 288-379, wherein the DNMT inhibitor comprises 5-Azacytidine.
The composition of claim 381, wherein 5-Azacytidine is present at about 0.2 μM to about 20 μM in the composition.
The composition of claim 381, wherein 5-Azacytidine is present at about 0.5 μM to about 10 μM in the composition.
The composition of claim 381, wherein 5-Azacytidine is present at about 1 μM to about 5 μM in the composition.
The composition of claim 381, wherein 5-Azacytidine is present at about 2 μM in the composition.
The composition of any one of claims 288-385, wherein the p38 MAPK inhibitor comprises BIRB796, SB203580, or SB202190.
The composition of any one of claims 288-385, wherein the p38 MAPK inhibitor comprises BIRB796.
The composition of claim 387, wherein BIRB796 is present at about 0.2 μM to about 20 μM in the composition.
The composition of claim 387, wherein BIRB796 is present at about 0.4 μM to about 10 μM in the composition.
The composition of claim 387, wherein BIRB796 is present at about 0.8 μM to about 5 μM in the composition.
The composition of claim 387, wherein BIRB796 is present at about 2 μM in the composition.
The composition of any one of claims 288-391, wherein the composition is a medium for culturing cells in vitro.
A composition comprising:

(a) somatic cells, and

(b) a c-Jun kinase inhibitor or a CBP/p300 bromodomain inhibitor.
The composition of claim 393, wherein the composition further comprises a serine-threonine kinase Akt inhibitor.
A composition comprising:

(a) somatic cells, and

(b) one or more of a c-Jun kinase inhibitor, a CBP/p300 bromodomain inhibitor, or a serine-threonine kinase Akt inhibitor.
The composition of any one of claims 393-395, wherein the composition further comprises a Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, a Menin-MLL interaction inhibitor, or both.
The composition of any one of claims 393-396, wherein the composition further comprises one or more of:

(1) a glycogen kinase inhibitor;

(2) a TGFβ receptor inhibitor;

(3) a retinoic acid receptor agonist;

(4) an agonist for the G protein-coupled receptor Smoothened;

(5) a Dot1L inhibitor;

(6) a Jak1/Jak2 inhibitor; or

(7) an SAH hydrolase inhibitor.
The composition of any one of claims 393-397, wherein the composition further comprises one or more of an adenosine kinase inhibitor, a casein kinase 2 inhibitor, a G9a inhibitor, a DNMT inhibitor, a p38 MAPK inhibitor.
The composition of any one of claims 393-397, wherein the composition further comprises a G9a inhibitor or a DNMT inhibitor.
The composition of claim 398, wherein the composition comprises the adenosine kinase inhibitor, the casein kinase 2 inhibitor, or both.
The composition of any one of claims 393-400, wherein the somatic cells comprise primary human adult adipose derived mesenchymal stromal cells (hADSCs) .
The composition of any one of claims 393-400, wherein the somatic cells comprise fibroblasts.
The composition of any one of claims 393-402, wherein the somatic cells comprise a genetic modification.
The composition of claim 403, wherein the genetic modification comprises an exogenous nucleic acid sequence.
The composition of claim 404, wherein the exogenous nucleic acid sequence encodes a polypeptide.
The composition of claim 404, wherein the exogenous nucleic acid sequence comprises a sequence of a non-coding nucleic acid molecule.
The composition of any one of claims 403-406, wherein the genetic modification comprises alteration of a genomic sequence.
The composition of any one of claims 403-407, wherein the genetic modification reduces immunogenicity of the somatic cells.