JP5560391B2 - ヒト胚性幹細胞の懸濁培養法 - Google Patents
ヒト胚性幹細胞の懸濁培養法 Download PDFInfo
- Publication number
- JP5560391B2 JP5560391B2 JP2008518312A JP2008518312A JP5560391B2 JP 5560391 B2 JP5560391 B2 JP 5560391B2 JP 2008518312 A JP2008518312 A JP 2008518312A JP 2008518312 A JP2008518312 A JP 2008518312A JP 5560391 B2 JP5560391 B2 JP 5560391B2
- Authority
- JP
- Japan
- Prior art keywords
- cells
- medium
- culture
- hes
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000004114 suspension culture Methods 0.000 title claims description 32
- 238000000034 method Methods 0.000 title claims description 30
- 210000001671 embryonic stem cell Anatomy 0.000 title description 13
- 210000004027 cell Anatomy 0.000 claims description 319
- 239000002609 medium Substances 0.000 claims description 69
- 238000012258 culturing Methods 0.000 claims description 21
- 239000000725 suspension Substances 0.000 claims description 17
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims description 15
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims description 15
- 210000002744 extracellular matrix Anatomy 0.000 claims description 15
- 210000000130 stem cell Anatomy 0.000 claims description 15
- 239000007787 solid Substances 0.000 claims description 14
- 108010085895 Laminin Proteins 0.000 claims description 13
- 102000007547 Laminin Human genes 0.000 claims description 13
- 239000001963 growth medium Substances 0.000 claims description 11
- 235000015097 nutrients Nutrition 0.000 claims description 11
- 102100020715 Fms-related tyrosine kinase 3 ligand protein Human genes 0.000 claims description 7
- 101710162577 Fms-related tyrosine kinase 3 ligand protein Proteins 0.000 claims description 7
- 102000018233 Fibroblast Growth Factor Human genes 0.000 claims description 5
- 108050007372 Fibroblast Growth Factor Proteins 0.000 claims description 5
- 229940126864 fibroblast growth factor Drugs 0.000 claims description 5
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims description 4
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims description 4
- 239000011859 microparticle Substances 0.000 claims description 4
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 claims description 3
- 101001027128 Homo sapiens Fibronectin Proteins 0.000 claims 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 30
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 30
- 239000003636 conditioned culture medium Substances 0.000 description 25
- 230000004069 differentiation Effects 0.000 description 24
- 230000012010 growth Effects 0.000 description 19
- 230000014509 gene expression Effects 0.000 description 17
- 210000001519 tissue Anatomy 0.000 description 16
- 210000001654 germ layer Anatomy 0.000 description 14
- 101150021185 FGF gene Proteins 0.000 description 13
- 210000001778 pluripotent stem cell Anatomy 0.000 description 12
- 238000004113 cell culture Methods 0.000 description 11
- 108010082117 matrigel Proteins 0.000 description 10
- 241000288906 Primates Species 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 210000001900 endoderm Anatomy 0.000 description 8
- 239000003550 marker Substances 0.000 description 8
- 239000003102 growth factor Substances 0.000 description 7
- 239000002245 particle Substances 0.000 description 7
- 230000035755 proliferation Effects 0.000 description 7
- 101000655352 Homo sapiens Telomerase reverse transcriptase Proteins 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 210000003981 ectoderm Anatomy 0.000 description 6
- 210000002950 fibroblast Anatomy 0.000 description 6
- 239000012737 fresh medium Substances 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000011159 matrix material Substances 0.000 description 6
- 210000003716 mesoderm Anatomy 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 206010043276 Teratoma Diseases 0.000 description 5
- 210000002459 blastocyst Anatomy 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000001143 conditioned effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000002242 embryoid body Anatomy 0.000 description 5
- 239000003797 essential amino acid Substances 0.000 description 5
- 235000020776 essential amino acid Nutrition 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 210000003494 hepatocyte Anatomy 0.000 description 5
- 238000003365 immunocytochemistry Methods 0.000 description 5
- 239000003226 mitogen Substances 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108020003175 receptors Proteins 0.000 description 5
- 102000005962 receptors Human genes 0.000 description 5
- 102000029816 Collagenase Human genes 0.000 description 4
- 108060005980 Collagenase Proteins 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 229960002424 collagenase Drugs 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000000877 morphologic effect Effects 0.000 description 4
- 238000010899 nucleation Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000000717 retained effect Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 4
- 102000007469 Actins Human genes 0.000 description 3
- 108010085238 Actins Proteins 0.000 description 3
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 description 3
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 description 3
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102100037362 Fibronectin Human genes 0.000 description 3
- 108010067306 Fibronectins Proteins 0.000 description 3
- 101001094700 Homo sapiens POU domain, class 5, transcription factor 1 Proteins 0.000 description 3
- 102100020880 Kit ligand Human genes 0.000 description 3
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 3
- 102000004058 Leukemia inhibitory factor Human genes 0.000 description 3
- 108090000581 Leukemia inhibitory factor Proteins 0.000 description 3
- 108010025020 Nerve Growth Factor Proteins 0.000 description 3
- 102000007072 Nerve Growth Factors Human genes 0.000 description 3
- 238000011579 SCID mouse model Methods 0.000 description 3
- 108010039445 Stem Cell Factor Proteins 0.000 description 3
- 238000013019 agitation Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229940112869 bone morphogenetic protein Drugs 0.000 description 3
- 210000004413 cardiac myocyte Anatomy 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012136 culture method Methods 0.000 description 3
- 210000003754 fetus Anatomy 0.000 description 3
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 210000004248 oligodendroglia Anatomy 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 108010049955 Bone Morphogenetic Protein 4 Proteins 0.000 description 2
- 102100024505 Bone morphogenetic protein 4 Human genes 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- OHCQJHSOBUTRHG-KGGHGJDLSA-N FORSKOLIN Chemical compound O=C([C@@]12O)C[C@](C)(C=C)O[C@]1(C)[C@@H](OC(=O)C)[C@@H](O)[C@@H]1[C@]2(C)[C@@H](O)CCC1(C)C OHCQJHSOBUTRHG-KGGHGJDLSA-N 0.000 description 2
- 102000004862 Gastrin releasing peptide Human genes 0.000 description 2
- 108090001053 Gastrin releasing peptide Proteins 0.000 description 2
- 102000003745 Hepatocyte Growth Factor Human genes 0.000 description 2
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 2
- 102000003964 Histone deacetylase Human genes 0.000 description 2
- 108090000353 Histone deacetylase Proteins 0.000 description 2
- 101000595198 Homo sapiens Podocalyxin Proteins 0.000 description 2
- 108091006905 Human Serum Albumin Proteins 0.000 description 2
- 102000008100 Human Serum Albumin Human genes 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- 101100335081 Mus musculus Flt3 gene Proteins 0.000 description 2
- 102000005604 Myosin Heavy Chains Human genes 0.000 description 2
- 108010084498 Myosin Heavy Chains Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108010068425 Octamer Transcription Factor-3 Proteins 0.000 description 2
- 102000002584 Octamer Transcription Factor-3 Human genes 0.000 description 2
- 102000004140 Oncostatin M Human genes 0.000 description 2
- 108090000630 Oncostatin M Proteins 0.000 description 2
- -1 PDGF Proteins 0.000 description 2
- 108010067787 Proteoglycans Proteins 0.000 description 2
- 102000016611 Proteoglycans Human genes 0.000 description 2
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 2
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 2
- 108010031318 Vitronectin Proteins 0.000 description 2
- 102100035140 Vitronectin Human genes 0.000 description 2
- 108010023082 activin A Proteins 0.000 description 2
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000005557 antagonist Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000007640 basal medium Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000007877 drug screening Methods 0.000 description 2
- 210000002308 embryonic cell Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- PUBCCFNQJQKCNC-XKNFJVFFSA-N gastrin-releasingpeptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)C(C)C)[C@@H](C)O)C(C)C)[C@@H](C)O)C(C)C)C1=CNC=N1 PUBCCFNQJQKCNC-XKNFJVFFSA-N 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 230000003394 haemopoietic effect Effects 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 108010055896 polyornithine Proteins 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 229930002330 retinoic acid Natural products 0.000 description 2
- 229910052711 selenium Inorganic materials 0.000 description 2
- 239000011669 selenium Substances 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 229960001727 tretinoin Drugs 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- LAQPKDLYOBZWBT-NYLDSJSYSA-N (2s,4s,5r,6r)-5-acetamido-2-{[(2s,3r,4s,5s,6r)-2-{[(2r,3r,4r,5r)-5-acetamido-1,2-dihydroxy-6-oxo-4-{[(2s,3s,4r,5s,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}hexan-3-yl]oxy}-3,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy}-4-hydroxy-6-[(1r,2r)-1,2,3-trihydrox Chemical compound O[C@H]1[C@H](O)[C@H](O)[C@H](C)O[C@H]1O[C@H]([C@@H](NC(C)=O)C=O)[C@@H]([C@H](O)CO)O[C@H]1[C@H](O)[C@@H](O[C@]2(O[C@H]([C@H](NC(C)=O)[C@@H](O)C2)[C@H](O)[C@H](O)CO)C(O)=O)[C@@H](O)[C@@H](CO)O1 LAQPKDLYOBZWBT-NYLDSJSYSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 101150033839 4 gene Proteins 0.000 description 1
- NMUSYJAQQFHJEW-UHFFFAOYSA-N 5-Azacytidine Natural products O=C1N=C(N)N=CN1C1C(O)C(O)C(CO)O1 NMUSYJAQQFHJEW-UHFFFAOYSA-N 0.000 description 1
- NMUSYJAQQFHJEW-KVTDHHQDSA-N 5-azacytidine Chemical compound O=C1N=C(N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 NMUSYJAQQFHJEW-KVTDHHQDSA-N 0.000 description 1
- HFDKKNHCYWNNNQ-YOGANYHLSA-N 75976-10-2 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](C)N)C(C)C)[C@@H](C)O)C1=CC=C(O)C=C1 HFDKKNHCYWNNNQ-YOGANYHLSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 1
- 108010002913 Asialoglycoproteins Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- 108010049931 Bone Morphogenetic Protein 2 Proteins 0.000 description 1
- 102100024506 Bone morphogenetic protein 2 Human genes 0.000 description 1
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 1
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 1
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 1
- 108010005939 Ciliary Neurotrophic Factor Proteins 0.000 description 1
- 102100031614 Ciliary neurotrophic factor Human genes 0.000 description 1
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 description 1
- 102000002004 Cytochrome P-450 Enzyme System Human genes 0.000 description 1
- 108010015742 Cytochrome P-450 Enzyme System Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 230000007067 DNA methylation Effects 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- SUZLHDUTVMZSEV-UHFFFAOYSA-N Deoxycoleonol Natural products C12C(=O)CC(C)(C=C)OC2(C)C(OC(=O)C)C(O)C2C1(C)C(O)CCC2(C)C SUZLHDUTVMZSEV-UHFFFAOYSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 101001004391 Drosophila melanogaster Protein jim lovell Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102400001368 Epidermal growth factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 108091008794 FGF receptors Proteins 0.000 description 1
- 102000044168 Fibroblast Growth Factor Receptor Human genes 0.000 description 1
- 102100028072 Fibroblast growth factor 4 Human genes 0.000 description 1
- 108090000381 Fibroblast growth factor 4 Proteins 0.000 description 1
- 102400000321 Glucagon Human genes 0.000 description 1
- 108060003199 Glucagon Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000003638 Glucose-6-Phosphatase Human genes 0.000 description 1
- 108010086800 Glucose-6-Phosphatase Proteins 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- 108010090007 Homeobox Protein Nkx-2.5 Proteins 0.000 description 1
- 102000012808 Homeobox Protein Nkx-2.5 Human genes 0.000 description 1
- 101000976075 Homo sapiens Insulin Proteins 0.000 description 1
- 101001092197 Homo sapiens RNA binding protein fox-1 homolog 3 Proteins 0.000 description 1
- 101000766306 Homo sapiens Serotransferrin Proteins 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 241000282560 Macaca mulatta Species 0.000 description 1
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000008730 Nestin Human genes 0.000 description 1
- 108010088225 Nestin Proteins 0.000 description 1
- 108010069196 Neural Cell Adhesion Molecules Proteins 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 102100037369 Nidogen-1 Human genes 0.000 description 1
- 102000016979 Other receptors Human genes 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000018886 Pancreatic Polypeptide Human genes 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 102000001393 Platelet-Derived Growth Factor alpha Receptor Human genes 0.000 description 1
- 108010068588 Platelet-Derived Growth Factor alpha Receptor Proteins 0.000 description 1
- 102100036031 Podocalyxin Human genes 0.000 description 1
- 102000003923 Protein Kinase C Human genes 0.000 description 1
- 108090000315 Protein Kinase C Proteins 0.000 description 1
- 108010014608 Proto-Oncogene Proteins c-kit Proteins 0.000 description 1
- 102000016971 Proto-Oncogene Proteins c-kit Human genes 0.000 description 1
- 101710183548 Pyridoxal 5'-phosphate synthase subunit PdxS Proteins 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- 102100035530 RNA binding protein fox-1 homolog 3 Human genes 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 101100537532 Rattus norvegicus Tnni3 gene Proteins 0.000 description 1
- 101150086694 SLC22A3 gene Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 102000005157 Somatostatin Human genes 0.000 description 1
- 108010056088 Somatostatin Proteins 0.000 description 1
- 101000983124 Sus scrofa Pancreatic prohormone precursor Proteins 0.000 description 1
- 108091005735 TGF-beta receptors Proteins 0.000 description 1
- 108010017842 Telomerase Proteins 0.000 description 1
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical compound IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 102000016715 Transforming Growth Factor beta Receptors Human genes 0.000 description 1
- 108010009583 Transforming Growth Factors Proteins 0.000 description 1
- 102000009618 Transforming Growth Factors Human genes 0.000 description 1
- 102100026893 Troponin T, cardiac muscle Human genes 0.000 description 1
- 101710165323 Troponin T, cardiac muscle Proteins 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 108091000117 Tyrosine 3-Monooxygenase Proteins 0.000 description 1
- 102000048218 Tyrosine 3-monooxygenases Human genes 0.000 description 1
- IVOMOUWHDPKRLL-UHFFFAOYSA-N UNPD107823 Natural products O1C2COP(O)(=O)OC2C(O)C1N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-UHFFFAOYSA-N 0.000 description 1
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- VREFGVBLTWBCJP-UHFFFAOYSA-N alprazolam Chemical compound C12=CC(Cl)=CC=C2N2C(C)=NN=C2CN=C1C1=CC=CC=C1 VREFGVBLTWBCJP-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000002744 anti-aggregatory effect Effects 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960002756 azacitidine Drugs 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 208000036815 beta tubulin Diseases 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000002798 bone marrow cell Anatomy 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 108010039524 chondroitin sulfate proteoglycan 4 Proteins 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000010372 cloning stem cell Methods 0.000 description 1
- 239000007376 cm-medium Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- OHCQJHSOBUTRHG-UHFFFAOYSA-N colforsin Natural products OC12C(=O)CC(C)(C=C)OC1(C)C(OC(=O)C)C(O)C1C2(C)C(O)CCC1(C)C OHCQJHSOBUTRHG-UHFFFAOYSA-N 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 229940095074 cyclic amp Drugs 0.000 description 1
- 230000002559 cytogenic effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 210000005064 dopaminergic neuron Anatomy 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 238000013100 final test Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 108700014844 flt3 ligand Proteins 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 1
- 229960004666 glucagon Drugs 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 208000013210 hematogenous Diseases 0.000 description 1
- 229940121372 histone deacetylase inhibitor Drugs 0.000 description 1
- 239000003276 histone deacetylase inhibitor Substances 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 238000012760 immunocytochemical staining Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 description 1
- 210000004153 islets of langerhan Anatomy 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- 108010028309 kalinin Proteins 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 241001515942 marmosets Species 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000000921 morphogenic effect Effects 0.000 description 1
- 210000005055 nestin Anatomy 0.000 description 1
- 210000000276 neural tube Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 108010008217 nidogen Proteins 0.000 description 1
- 102000045246 noggin Human genes 0.000 description 1
- 108700007229 noggin Proteins 0.000 description 1
- 230000006911 nucleation Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000001582 osteoblastic effect Effects 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 229940021222 peritoneal dialysis isotonic solution Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 1
- 229920000768 polyamine Polymers 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920002714 polyornithine Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011165 process development Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- XNSAINXGIQZQOO-SRVKXCTJSA-N protirelin Chemical compound NC(=O)[C@@H]1CCCN1C(=O)[C@@H](NC(=O)[C@H]1NC(=O)CC1)CC1=CN=CN1 XNSAINXGIQZQOO-SRVKXCTJSA-N 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 210000005084 renal tissue Anatomy 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000012340 reverse transcriptase PCR Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229940091258 selenium supplement Drugs 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000002536 stromal cell Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 229940035722 triiodothyronine Drugs 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 102000009310 vitamin D receptors Human genes 0.000 description 1
- 108050000156 vitamin D receptors Proteins 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 210000001325 yolk sac Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/113—Acidic fibroblast growth factor (aFGF, FGF-1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/119—Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/15—Transforming growth factor beta (TGF-β)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/26—Flt-3 ligand (CD135L, flk-2 ligand)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/52—Fibronectin; Laminin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Reproductive Health (AREA)
- Developmental Biology & Embryology (AREA)
- Gynecology & Obstetrics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
本出願は、2005年6月22日に出願された米国特許出願第60/693266号の優先権を主張する。
再生医療は、様々な種類の始原細胞の分離、培養、利用に関連する最近の進歩によって恩恵を受けている。この発明の開示は、ヒト多能性幹細胞とその誘導体の商業的開発の更なる進歩を与えるものである。
本開示は、霊長類多能性幹細胞(hES細胞)の培養および増殖のための改良されたシステムを提供するものである。この発明に係る懸濁培養システムは、治療及び創薬における使用のために、早くかつ容量効率的に高品質の胚性幹細胞を生産させることができる。
霊長類の多能性幹(hES)細胞細胞を増殖させる従来の技術は、固体表面上での培養を含むものであった。即ち、繊維芽細胞フィーダー細胞法(米国特許第6,200,806号)、または細胞外マトリックス法(米国特許第6,800,480号)であった。フィーダーフリーの方法は、迅速な増幅を可能にするように最適化されるのでWO 03/020920、商業目的のhES細胞製造コストを大幅に減少させる。
始原型である「霊長類多能性幹細胞」(pPS細胞)は、受精後の任意の時期における前胚組織、胚組織、または胎児組織に由来する多能性細胞であり、正常な条件の下でいくつかの異なる細胞型の子孫を作ることが可能であるという特徴を有する。pPS細胞は、適当な宿主において奇形腫を形成する能力、または培養液において三つの胚葉全ての組織型のマーカーを持つ細胞に分化する能力のように、標準的な技術的に容認された試験に従って、三つの胚葉、即ち内胚葉、中胚葉、および外胚葉のそれぞれの誘導体である子孫を作ることができる。
分子遺伝学、並びに遺伝子工学の一般的方法は、Molecular Cloning: A Laboratory Manual, (Sambrook et al., Cold Spring Harbor);Gene Transfer Vectors for Mammalian Cells (Miller & Calos eds.);およびCurrent Protocols in Molecular Biology (F. M. Ausubel et al. eds., Wiley & Sons) の最新版に記載されている。細胞生物学、たんぱく質化学、および抗体技術は、Current Protocols in Protein Science (J. E. Colligan et al. eds., Wiley & Sons); Current Protocols in Cell Biology (J. S. Bonifacino et al., Wiley & Sons)、 およびCurrent Protocols in Immunology (J. E. Colligan et al. eds., Wiley & Sons)に見ることができる。この開示において引用される試薬、クローニングベクター、および遺伝子操作のためのキット類は、Biorad、Stratagene、Invitrogen、Clontech、およびSigma-Aldrich Co.等の民間の供給業者から入手可能である。
胚性幹細胞は、霊長類種のメンバーの胚盤胞から採取することができる(米国特許第5,843,780号; Thomson et al., Proc. Natl. Acad. Sci. USA 92:7844, 1995)。ヒト胚性幹(hES)細胞は、Thomson et al. (米国特許第6,200,806号; Science 282:1145, 1998; Curr. Top. Dev. Biol. 38:133 ff., 1998)およびReubinoff et al. (Nature Biotech. 18:399, 2000)の技術に従って、初代マウス繊維芽細胞フィーダー細胞を用いてヒト胚盤胞細胞から調製することができる。hES細胞株はまた、ヒトフィーダー細胞上で(米国特許第6,642,048号)、または完全にフィーダーフリーの条件で(US 2002/0081724 A1)誘導されうる。hES細胞と同等の細胞型は、WO 01/51610 (Bresagen) において概説されたように、原始外胚葉様(EPL)細胞のような多能性派生物を含む。
最初にThomson (U.S. 5,843,780; U.S. 6,090,622)によって記載されたように、当初、この分野の科学者の多くは、分化を防ぐ目的でフィーダー細胞層の上でhES細胞を培養する方法を選んだ。
現在では、hES細胞は、固体の基質上で増殖させるよりもむしろ、懸濁培養によって増殖させることができることが発見されている。
・幹細胞因子(SCF, Steel factor)、c-kitを二量体化する他のリガンドまたは抗体、および同じシグナル伝達経路の他の活性化因子
・他のチロシンキナーゼ関連受容体に対するリガンド、例えば、血小板由来成長因子(PDGF), マクロファージコロニー刺激因子、Flt-3リガンド、および血管内皮成長因子(VEGF)の受容体
・サイクリックAMPレベルを上昇させる因子、例えば、フォルスコリン
・gp130を誘導する因子、例えば、LIFまたはオンコスタチン-M
・造血成長因子、例えば、トロンボポイチン(TPO)
・トランスフォーミング成長因子、例えば、TGFβ1
・他の成長因子、例えば上皮成長因子(EGF)
・ニューロトロフィン、例えばCNTF
この発明に従って培養されたヒトES細胞は、未分化幹細胞に特徴的な形態学的性質を持つ。二次元の標準的な顕微鏡像では、hES細胞は、画像の平面における高い核/細胞質比、よく目立つ核小体、およびよく識別できない細胞境界部を持つ密集したコロニー形成を有する。細胞株は、標準のGバンド法を用いて核型が調べられ(Oakland, CAのCytogenetics Labなどの日常業務の核型決定サービス業務を行う多くの臨床診断ラボが利用可能である)、公表されたヒト核型と比較される。細胞が正倍数体であり、顕著な変化をしていない全てのヒト染色体を持つ、「正常な核型」を持つ細胞を入手することが望ましい。
本発明は、多数の多能性細胞を商業的規模で生産する方法を提供するものである。該細胞は、未分化の状態で多数の研究および商業的目的で役に立つものであり、または、特定の細胞型へ分化させることに向けることができる。
肝細胞は、米国特許第6,458,589号およびPCT公報 WO 01/81549 (Geron Corporation) に記載されたように、ヒストンデアセチラーゼの阻害剤を用いてhES細胞から分化させることができる。未分化のhES細胞は、ヒストンデアセチラーゼの阻害剤の存在下で培養される。
神経細胞は、米国特許第6,833,269号; Carpentar et al., Exp. Neurol. 2001; 172(2):383-97; およびWO 03/000868 (Geron Corporation)に記載された方法に従ってhES細胞から生成されることができる。未分化hES細胞または胚様体細胞を、一つまたは複数のニューロトロフィン、および一つまたは複数の分裂促進物質を含有する培地で培養し、それにより少なくとも約60%の細胞がA2B5、ポリシアル酸化NCAM、またはネスチンを発現する細胞集団を生成し、かつこれは培養中に少なくとも20回の倍化を行うことができる。分裂促進物質の例は、EGF、塩基性FGF、PDGF、およびIGF-1である。ニューロトロフィンの例としては、NT-3およびBDNFがある。TGF-βスーパーファミリー拮抗剤の利用、またはcAMPとアスコルビン酸の組み合わせは、ドーパミン作動性ニューロンに特徴的なチロシンヒドロキシラーゼ陽性の神経細胞の比率を増加させるために使用することができる。増殖細胞は、さらに、分裂促進物質のない状態でニュートロフィンと共に培養することによって最終分化を行わせることができる。
心筋細胞または心筋細胞前駆体は、WO 03/006950に記載された方法によってhES細胞から生成させることができる。細胞は、ウシ胎児血清または血清代替物、および任意で5-アザシチジンなどのDNAのメチル化に影響する心臓の栄養因子とともに懸濁状態で培養される。また、心筋細胞の集団は、アクチビンAおよび骨形態形成たんぱく質4の組み合わせを用いて固体基質上で直接分化させることができる:かくして、収縮細胞を、密度遠心分離によって集団中の他の細胞から分離することができる。
膵島細胞は、アクチビンA、ヒストンデアセチラーゼ阻害剤(酪酸など)、分裂促進剤(bFGFなど)、およびTGF-βスーパーファミリー拮抗剤(ノギンなど)から選択されるいくつかの因子の組み合わせを含有する培地で培養することによってhES細胞の分化を開始させることにより、hES細胞から分化され得る。その後、該細胞をニコチンアミドと培養することによって成熟させることができ、細胞の少なくとも5%がPdx1、インスリン、グルカゴン、ソマトスタチン、および膵臓ポリペプチドを発現する細胞集団が得られる。WO 03/050249 (Geron Corp.)を参照されたい。
この発明に係る培養系の構成品は、売り物として提供され、販売され、またはそうでなければ、任意の目的での任意の企業による使用のために、製造場所から配布される。また、構成物は、次の二つまたはそれ以上の組み合わせの様々な有用な組み合わせで、一緒に販売または配布され得る。
・hES細胞を懸濁因子中で培養するための適当な培地
・培地に存在する、または添加されるべき細胞外マトリックス成分または増粘剤
・培地に存在する、または添加されるべきマイクロキャリア
・懸濁培養に合わせて改良された容器
・培養系において増殖している、または他の形態で貯蔵されるが培養系での使用が意図されたhES細胞それ自身
実施例1:迅速増殖培地における多能性幹細胞の成長
最初にマウス胚繊維芽細胞のフィーダー上で成長させ、それから米国特許第6,800,480号; Xu et al., Stem Cells 2005;23(3):315-23に詳しく述べられたようにMatrigel(商標登録)細胞外マトリックスと馴化培地を含むフィーダーフリー環境で20継代の間増殖させた、一系統のhES細胞を得た。
Matrigel(商標登録)上でMEF-CMにおいて培養されたhES細胞は、新鮮な(非馴化)血清非含有培地:グルタミン、非必須アミノ酸、およびβ-メルカプトエタノール+ Matrigel(商標登録)上の80 ng/mLヒト塩基性FGFで補足され、さらにヒトラミニンでコーティングされた表面に適合されたX-VIVO(商標)で継代された。あるいは、低温保存された細胞が、80 ng/mL hbFGFを含有する同じ培地に直接融解された。細胞は、コラゲナーゼIVを用いて5〜6日毎に継代された。
培養液の体積当りのhES細胞の収量を増やす目的で、細胞は懸濁培養され、次に、形態および三つ全ての胚葉を代表する分化した細胞になる能力が調べられた。
・容器:100 mLスピナーフラスコ
・接種(播種)密度:3.6 × 105 cells/mL
・培地量:スピナーフラスコ当り50 ml
・使用された培地:bFGF (8 ng/mL)を含有するmEF馴化培地
・攪拌速度:20 rpm (Bellco・キャリアー・マグネチック・スターラー)
・環境:37℃ CO2インキュベーター
・培地交換:隔日(細胞を沈下させ、上清を置換する)
H9 hES細胞は、これらの条件下で6日間スピナーフラスコ中で維持された。
・容器:100 mLスピナーフラスコ
・接種(播種)密度:3.5 × 105 細胞/mL
・培地量:スピナーフラスコ当り50 mL
・使用された培地: mEF-CM + bFGF (8 ng/mL)
・攪拌速度:20 rpm (前述)
・培地交換:三日に一度
この実験は、別のhES細胞株を用いて行われた。細胞は、いくつかの異なる培養条件下でスピナーフラスコの代わりにシェーカーフラスコを用いて培養され、培養は、2ヶ月以上継続された。
・容器:100 mLシェーカーフラスコ
・接種(播種)密度:5.0 × 105 細胞/mL
・培地量:シェーカーフラスコ当り15 ml
・攪拌速度:80 rpm (37℃ CO2インキュベーター中のLabline ローテータ/シェーカー)
・培地交換:最初は隔日、その後2〜3日毎
使用された培地および培養期間は次の通りである。
A: mEF-CM + bFGF (8 ng/mL)。98日間維持。
B: mEF-CM + bFGF (8 ng/mL) + ラミニン(最初に33 μg/mL、その後の残りの培養で約10μg/ml)。49日間維持。
C: X-VIVO(商標)10 + FGF (40 ng/mL) + Flt-3 (75 ng/mL)。11日間維持。
D: X-VIVO(商標) 10 + FGF (40 ng/mL) + Flt-3 (75 ng/mL) + ラミニン(最初に33 μg/mL、その後の残りの培養で約10μg/ml)。11日間維持。
培養期間中の細胞数は次の表に示される。
(表3)懸濁培養におけるhES細胞の増殖
次の実験では、懸濁培養において新鮮(非馴化)培地を用いる場合の代替えの添加物を評価する。
1) X-VIVO(商標) 10+ bFGF (80ng/mL)
2) X-VIVO(商標) 10+ bFGF (80ng/mL) + TGFβ1 (0.5 ng/mL)
3) X-VIVO(商標) 10+ bFGF (40ng/mL) + TGFβ1 (0.5 ng/mL)
4) X-VIVO(商標) 10+ bFGF (80ng/mL) + TGFβ1 (0.5 ng/mL) + 10μg/mLヒトラミニン
5) X-VIVO 10(商標)+ bFGF (80ng/mL) + TGFβ1 (0.5 ng/mL) + 50μg/mL ヒト血清アルブミン
Claims (11)
- hES細胞を実質的に未分化の状態に維持しながら培養する方法であって、以下:
a)該細胞を栄養培地に懸濁し;
b)培養している間、該細胞を懸濁状態に保ち;
c)培地を定期的に交換し;
d)必要に応じて培養液を時々分割し、細胞密度を減少させることによりhES細胞を実質的に未分化の状態で培養する工程を含む方法。 - 細胞が少なくとも約40ng/mLの濃度の繊維芽細胞成長因子を含有する培地で培養される、請求項1に記載の方法。
- 培地がトランスフォーミング成長因子ベータ(TGFβ)、幹細胞因子(SCF)、またはFlt3リガンド (Flt3L)も含有する、請求項2に記載の方法。
- 細胞が一つまたは複数の、可溶性のまたは懸濁された細胞外マトリックス成分を含有する培地で培養される、請求項1から3のいずれか一項に記載の方法。
- 細胞外マトリックス成分がヒトラミニンおよび/またはヒトフィブロネクチンを含む、請求項4に記載の方法。
- 細胞が固体微粒子と共に懸濁培養される、請求項1から5のいずれか一項に記載の方法。
- 微粒子が、一つまたは複数の、可溶性のまたは懸濁された細胞外マトリックス成分でコーティングされる、請求項6に記載の方法。
- 培養されたhES細胞を収集し、実質的に未分化の細胞集団が維持されるように、収集した細胞を固体表面にプレーティングして、該細胞の培養を継続する工程を更に含む、請求項1から7のいずれか一項に記載の方法。
- 培養されたhES細胞を収集し、収集された細胞を分化させる工程を更に含む、請求項1から8のいずれか一項に記載の方法。
- 少なくとも約40ng/mLの繊維芽細胞成長因子を含有する栄養培地を含む、hES細胞を懸濁培養するためのシステムまたはキット。
- 培地に添加するための細胞外マトリックス成分または微粒子も含む、請求項10に記載のシステムまたはキット。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US69326605P | 2005-06-22 | 2005-06-22 | |
US60/693,266 | 2005-06-22 | ||
PCT/US2006/023976 WO2007002086A2 (en) | 2005-06-22 | 2006-06-20 | Suspension culture of human embryonic stem cells |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013035335A Division JP5770213B2 (ja) | 2005-06-22 | 2013-02-26 | ヒト胚性幹細胞の懸濁培養法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2008546410A JP2008546410A (ja) | 2008-12-25 |
JP5560391B2 true JP5560391B2 (ja) | 2014-07-23 |
Family
ID=37595746
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008518312A Active JP5560391B2 (ja) | 2005-06-22 | 2006-06-20 | ヒト胚性幹細胞の懸濁培養法 |
JP2013035335A Active JP5770213B2 (ja) | 2005-06-22 | 2013-02-26 | ヒト胚性幹細胞の懸濁培養法 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013035335A Active JP5770213B2 (ja) | 2005-06-22 | 2013-02-26 | ヒト胚性幹細胞の懸濁培養法 |
Country Status (11)
Country | Link |
---|---|
US (5) | US9074181B2 (ja) |
EP (2) | EP1910516B1 (ja) |
JP (2) | JP5560391B2 (ja) |
KR (4) | KR20150067394A (ja) |
CN (2) | CN107189980B (ja) |
AU (1) | AU2006262369B2 (ja) |
CA (1) | CA2613369C (ja) |
GB (1) | GB2441488C (ja) |
HK (1) | HK1122836A1 (ja) |
IL (1) | IL188264A (ja) |
WO (1) | WO2007002086A2 (ja) |
Families Citing this family (72)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8597947B2 (en) * | 2004-12-29 | 2013-12-03 | Hadasit Medical Research Services & Development Limited | Undifferentiated stem cell culture systems |
EP2410043A3 (en) | 2004-12-29 | 2013-01-23 | Hadasit Medical Research Services And Development Ltd. | Stem cells culture systems |
CN107189980B (zh) | 2005-06-22 | 2021-07-09 | 阿斯特利亚斯生物治疗股份公司 | 人胚胎干细胞的悬浮培养物 |
EP1962719A4 (en) * | 2005-08-29 | 2011-05-04 | Technion Res And Dev Of Foundation Ltd | MEDIA FOR BREEDING STEM CELLS |
AU2015203310B2 (en) * | 2005-08-29 | 2017-04-20 | Technion Research & Development Foundation Ltd. | Media for Culturing Stem Cells |
PT2733203T (pt) | 2006-08-02 | 2019-01-23 | Technion Res & Dev Foundation | Métodos de expansão de células estminais embrionárias numa cultura de suspensão |
US9080145B2 (en) | 2007-07-01 | 2015-07-14 | Lifescan Corporation | Single pluripotent stem cell culture |
KR101617243B1 (ko) | 2007-07-31 | 2016-05-02 | 라이프스캔, 인코포레이티드 | 인간 배아 줄기 세포의 분화 |
KR101592182B1 (ko) | 2007-11-27 | 2016-02-05 | 라이프스캔, 인코포레이티드 | 인간 배아 줄기 세포의 분화 |
WO2009072990A1 (en) | 2007-12-06 | 2009-06-11 | Agency For Science, Technology And Research | Method for extracellular matrix mediated differentiation and proliferation of stem cells |
KR101731474B1 (ko) | 2008-02-21 | 2017-05-11 | 얀센 바이오테크 인코포레이티드 | 세포 부착, 배양 및 탈리를 위한 방법, 표면 개질 플레이트 및 조성물 |
US8093053B2 (en) | 2008-03-17 | 2012-01-10 | Uti Limited Partnership | Methods and compositions for culturing of neural precursor cells |
US8716018B2 (en) * | 2008-03-17 | 2014-05-06 | Agency For Science, Technology And Research | Microcarriers for stem cell culture |
US9458431B2 (en) | 2008-03-17 | 2016-10-04 | Agency For Science, Technology And Research | Microcarriers for stem cell culture |
WO2009147400A1 (en) * | 2008-06-05 | 2009-12-10 | Iti Scotland Limited | Stem cell culture media and methods |
PL2942392T3 (pl) | 2008-06-30 | 2019-02-28 | Janssen Biotech, Inc | Różnicowanie pluripotencjalnych komórek macierzystych |
US20100120142A1 (en) | 2008-07-11 | 2010-05-13 | Suomen Punainen Risti Veripalvelu | Culture of human embryonic cells |
CN102333862B (zh) | 2008-10-31 | 2018-04-27 | 詹森生物科技公司 | 人胚胎干细胞向胰腺内分泌谱系的分化 |
US9234178B2 (en) | 2008-10-31 | 2016-01-12 | Janssen Biotech, Inc. | Differentiation of human pluripotent stem cells |
US8895300B2 (en) | 2008-11-04 | 2014-11-25 | Viacyte, Inc. | Scalable primate pluripotent stem cell aggregate suspension culture and differentiation thereof |
JP5390624B2 (ja) * | 2008-11-04 | 2014-01-15 | バイアサイト インク | 幹細胞集合体懸濁液組成物、その分化方法 |
JP5719305B2 (ja) | 2008-11-20 | 2015-05-13 | ヤンセン バイオテツク,インコーポレーテツド | 平面支持体上での細胞付着及び培養のための方法及び組成物 |
AU2009316580B2 (en) | 2008-11-20 | 2016-04-14 | Janssen Biotech, Inc. | Pluripotent stem cell culture on micro-carriers |
WO2011011300A2 (en) | 2009-07-20 | 2011-01-27 | Centocor Ortho Biotech Inc. | Differentiation of human embryonic stem cells |
EP4166652A1 (en) * | 2009-11-12 | 2023-04-19 | Technion Research & Development Foundation Ltd. | Culture media, cell cultures and methods of culturing pluripotent stem cells in an undifferentiated state |
US9150833B2 (en) | 2009-12-23 | 2015-10-06 | Janssen Biotech, Inc. | Differentiation of human embryonic stem cells |
RU2607380C2 (ru) | 2010-03-01 | 2017-01-10 | Янссен Байотек, Инк. | Способы очистки клеток, производных от плюрипотентных стволовых клеток |
CA2828315A1 (en) | 2010-04-08 | 2011-10-13 | The University Court Of The University Of Edinburgh | Chondrogenic progenitor cells, protocol for derivation of cells and uses thereof |
WO2011143299A2 (en) | 2010-05-12 | 2011-11-17 | Centocor Ortho Biotech Inc. | Differentiation of human embryonic stem cells |
WO2012009377A2 (en) | 2010-07-12 | 2012-01-19 | University Of Southern California | Biocompatible substrate for facilitating interconnections between stem cells and target tissues and methods for implanting same |
EP3211070A1 (en) | 2010-08-31 | 2017-08-30 | Janssen Biotech, Inc. | Differentiation of human embryonic stem cells |
US9506036B2 (en) | 2010-08-31 | 2016-11-29 | Janssen Biotech, Inc. | Differentiation of human embryonic stem cells |
MX348537B (es) | 2010-08-31 | 2017-06-07 | Janssen Biotech Inc | Diferencia de celulas madre pluripotentes. |
CN102212460B (zh) * | 2011-04-27 | 2013-06-05 | 中国人民解放军第三军医大学第二附属医院 | 干细胞筛选系统、其制备方法及干细胞的筛选方法 |
US8877489B2 (en) | 2011-12-05 | 2014-11-04 | California Institute Of Technology | Ultrathin parylene-C semipermeable membranes for biomedical applications |
US10478206B2 (en) | 2011-04-29 | 2019-11-19 | University Of Southern California | Instruments and methods for the implantation of cell-seeded substrates |
EP2784153B1 (en) | 2011-11-25 | 2018-01-03 | Kyoto University | Method for culturing pluripotent stem cell |
US9248013B2 (en) | 2011-12-05 | 2016-02-02 | California Institute Of Technology | 3-Dimensional parylene scaffold cage |
RU2705001C2 (ru) | 2011-12-22 | 2019-11-01 | Янссен Байотек, Инк. | Дифференцировка эмбриональных стволовых клеток человека в одногормональные инсулинположительные клетки |
CN104160018A (zh) | 2012-03-07 | 2014-11-19 | 詹森生物科技公司 | 用于扩增和维持多能干细胞的成分确定的培养基 |
ES2690118T3 (es) | 2012-06-08 | 2018-11-19 | Janssen Biotech, Inc. | Diferenciación de células madre embrionarias humanas en células endocrinas pancreáticas |
EP3409761A1 (en) * | 2012-07-24 | 2018-12-05 | Nissan Chemical Corporation | Culture medium composition, and method for culturing cell or tissue using said composition |
USD1009775S1 (en) | 2014-10-15 | 2024-01-02 | Maxeon Solar Pte. Ltd. | Solar panel |
USD933584S1 (en) | 2012-11-08 | 2021-10-19 | Sunpower Corporation | Solar panel |
KR101942769B1 (ko) | 2012-12-31 | 2019-01-28 | 얀센 바이오테크 인코포레이티드 | Hb9 조절제를 사용하는 인간 배아 줄기세포의 췌장 내분비 세포로의 분화 |
CA2896750A1 (en) | 2012-12-31 | 2014-07-03 | Janssen Biotech, Inc. | Suspension and clustering of human pluripotent cells for differentiation into pancreatic endocrine cells |
US10370644B2 (en) | 2012-12-31 | 2019-08-06 | Janssen Biotech, Inc. | Method for making human pluripotent suspension cultures and cells derived therefrom |
BR112015015770A2 (pt) | 2012-12-31 | 2017-07-11 | Janssen Biotech Inc | cultivo de células-tronco embrionárias humanas na interface ar-líquido para diferenciação em células pancreáticas endócrinas |
JP6292415B2 (ja) | 2013-03-06 | 2018-03-14 | 国立大学法人京都大学 | 多能性幹細胞の培養システム及び多能性幹細胞の継代方法 |
CN106414718A (zh) | 2013-06-11 | 2017-02-15 | 哈佛学院校长同事会 | SC-β细胞以及用于产生其的组合物和方法 |
JP2014036660A (ja) * | 2013-10-09 | 2014-02-27 | Viacyte Inc | 幹細胞集合体懸濁液組成物、その分化方法 |
WO2015175307A1 (en) | 2014-05-16 | 2015-11-19 | Janssen Biotech, Inc. | Use of small molecules to enhance mafa expression in pancreatic endocrine cells |
USD999723S1 (en) | 2014-10-15 | 2023-09-26 | Sunpower Corporation | Solar panel |
USD913210S1 (en) | 2014-10-15 | 2021-03-16 | Sunpower Corporation | Solar panel |
USD933585S1 (en) | 2014-10-15 | 2021-10-19 | Sunpower Corporation | Solar panel |
USD896747S1 (en) | 2014-10-15 | 2020-09-22 | Sunpower Corporation | Solar panel |
WO2016088243A1 (ja) * | 2014-12-05 | 2016-06-09 | 株式会社ニコン | 判定装置、観察システム、観察方法、そのプログラム、細胞の製造方法、および細胞 |
CN107614678B (zh) | 2014-12-18 | 2021-04-30 | 哈佛学院校长同事会 | 干细胞来源的β细胞的产生方法及其使用方法 |
WO2016100909A1 (en) | 2014-12-18 | 2016-06-23 | President And Fellows Of Harvard College | METHODS FOR GENERATING STEM CELL-DERIVED β CELLS AND USES THEREOF |
US10443042B2 (en) | 2014-12-18 | 2019-10-15 | President And Fellows Of Harvard College | Serum-free in vitro directed differentiation protocol for generating stem cell-derived beta cells and uses thereof |
WO2016104541A1 (ja) * | 2014-12-24 | 2016-06-30 | 国立大学法人京都大学 | 内胚葉系細胞の製造方法、肝臓細胞の製造方法、膵臓細胞の製造方法、内胚葉系細胞の誘導促進剤、肝臓細胞の誘導促進キット、膵臓細胞の誘導促進キット、およびマイクロ流体デバイス |
US9944894B2 (en) | 2015-01-16 | 2018-04-17 | General Electric Company | Pluripotent stem cell expansion and passage using a rocking platform bioreactor |
MA45479A (fr) | 2016-04-14 | 2019-02-20 | Janssen Biotech Inc | Différenciation de cellules souches pluripotentes en cellules de l'endoderme de l'intestin moyen |
CA3081762A1 (en) | 2017-11-15 | 2019-05-23 | Semma Therapeutics, Inc. | Islet cell manufacturing compositions and methods of use |
JPWO2019181999A1 (ja) * | 2018-03-20 | 2021-02-04 | 富士フイルム株式会社 | 多能性幹細胞を三次元培養する方法 |
AU2019320072A1 (en) | 2018-08-10 | 2021-02-25 | Vertex Pharmaceuticals Incorporated | Stem cell derived islet differentiation |
CN110257332A (zh) * | 2019-07-08 | 2019-09-20 | 广东省赛莱拉干细胞研究院 | 一种诱导间充质干细胞分化成为多巴胺能神经元的培养基以及方法 |
JP2022158092A (ja) * | 2019-08-20 | 2022-10-17 | 昭和電工マテリアルズ株式会社 | 馴化細胞作製方法 |
US11001810B1 (en) | 2019-11-11 | 2021-05-11 | Lancell AB | Serum-free human pluripotent stem cell culture medium |
WO2021180781A1 (en) * | 2020-03-10 | 2021-09-16 | Life & Brain Gmbh | Up-scaled production of microglia-like/-precursor cells and macrophage cells using mesh macrocarriers |
CN111808807B (zh) * | 2020-07-21 | 2021-07-30 | 生物岛实验室 | 一种无需预先包被的间充质干细胞无血清培养基及其应用 |
CN113249303B (zh) * | 2021-05-07 | 2022-07-29 | 华中农业大学 | 血管内皮细胞生长因子在促进鸡原始生殖细胞增殖和迁移中的应用 |
Family Cites Families (61)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US264713A (en) | 1882-09-19 | Neck-strap buckle and rein-ring | ||
US17589A (en) | 1857-06-16 | Improvement in bending sheet-metal pans | ||
WO1990001541A1 (en) | 1988-08-04 | 1990-02-22 | Amrad Corporation Limited | In vitro propagation of embryonic stem cells |
US5332672A (en) | 1991-12-02 | 1994-07-26 | Regeneron Pharmaceuticals, Inc. | Prevention of ES cell differentiation by ciliary neurotrophic factor |
WO1994007997A1 (en) | 1992-10-06 | 1994-04-14 | The Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services | Long-term proliferation of primordial germ cells |
US7153684B1 (en) * | 1992-10-08 | 2006-12-26 | Vanderbilt University | Pluripotential embryonic stem cells and methods of making same |
US5453357A (en) | 1992-10-08 | 1995-09-26 | Vanderbilt University | Pluripotential embryonic stem cells and methods of making same |
GB9308271D0 (en) | 1993-04-21 | 1993-06-02 | Univ Edinburgh | Method of isolating and/or enriching and/or selectively propagating pluripotential animal cells and animals for use in said method |
US5405772A (en) | 1993-06-18 | 1995-04-11 | Amgen Inc. | Medium for long-term proliferation and development of cells |
US5583016A (en) | 1994-07-07 | 1996-12-10 | Geron Corporation | Mammalian telomerase |
US5914268A (en) | 1994-11-21 | 1999-06-22 | National Jewish Center For Immunology & Respiratory Medicine | Embryonic cell populations and methods to isolate such populations |
US5874301A (en) | 1994-11-21 | 1999-02-23 | National Jewish Center For Immunology And Respiratory Medicine | Embryonic cell populations and methods to isolate such populations |
US5843780A (en) | 1995-01-20 | 1998-12-01 | Wisconsin Alumni Research Foundation | Primate embryonic stem cells |
AU1119397A (en) | 1995-11-14 | 1997-06-05 | Regents Of The University Of Minnesota | Ex vivo culture of stem cells |
WO1997047734A1 (en) | 1996-06-14 | 1997-12-18 | The Regents Of The University Of California | In vitro derivation and culture of primate pluripotent stem cells and therapeutic uses thereof |
US6638763B1 (en) | 1997-01-07 | 2003-10-28 | University Of Tennessee Research Foundation | Isolated mammalian neural stem cells, methods of making such cells |
AU5734998A (en) | 1997-01-10 | 1998-08-03 | Life Technologies, Inc. | Embryonic stem cell serum replacement |
US6331406B1 (en) | 1997-03-31 | 2001-12-18 | The John Hopkins University School Of Medicine | Human enbryonic germ cell and methods of use |
US6090622A (en) | 1997-03-31 | 2000-07-18 | The Johns Hopkins School Of Medicine | Human embryonic pluripotent germ cells |
US5922567A (en) | 1997-06-03 | 1999-07-13 | Incyte Pharmaceuticals, Inc. | Two new human DNAJ-like proteins |
US5968829A (en) | 1997-09-05 | 1999-10-19 | Cytotherapeutics, Inc. | Human CNS neural stem cells |
US6800480B1 (en) | 1997-10-23 | 2004-10-05 | Geron Corporation | Methods and materials for the growth of primate-derived primordial stem cells in feeder-free culture |
WO1999042122A1 (en) | 1998-02-19 | 1999-08-26 | University Of Southern California | Method of promoting embryonic stem cell proliferation |
JP2002504362A (ja) | 1998-02-27 | 2002-02-12 | メディカル・カレッジ・オブ・ハンプトン・ローズ | 移植治療のための前駆胚幹細胞からの細胞および組織の誘導体 |
WO2000017323A1 (en) | 1998-09-22 | 2000-03-30 | Neuralstem Biopharmaceuticals, Ltd. | Stable neural stem cell lines |
US7410798B2 (en) | 2001-01-10 | 2008-08-12 | Geron Corporation | Culture system for rapid expansion of human embryonic stem cells |
US7413904B2 (en) | 1998-10-23 | 2008-08-19 | Geron Corporation | Human embryonic stem cells having genetic modifications |
US6667176B1 (en) | 2000-01-11 | 2003-12-23 | Geron Corporation | cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells |
IL142748A0 (en) | 1998-11-09 | 2002-03-10 | Univ Monash | Embryonic stem cells |
US6280718B1 (en) | 1999-11-08 | 2001-08-28 | Wisconsin Alumni Reasearch Foundation | Hematopoietic differentiation of human pluripotent embryonic stem cells |
US7455983B2 (en) * | 2000-01-11 | 2008-11-25 | Geron Corporation | Medium for growing human embryonic stem cells |
WO2001051610A1 (en) | 2000-01-14 | 2001-07-19 | Bresagen Limited | Ectoderm cell production |
US7005252B1 (en) | 2000-03-09 | 2006-02-28 | Wisconsin Alumni Research Foundation | Serum free cultivation of primate embryonic stem cells |
US6458589B1 (en) | 2000-04-27 | 2002-10-01 | Geron Corporation | Hepatocyte lineage cells derived from pluripotent stem cells |
US7473555B2 (en) | 2000-04-27 | 2009-01-06 | Geron Corporation | Protocols for making hepatocytes from embryonic stem cells |
US7250294B2 (en) | 2000-05-17 | 2007-07-31 | Geron Corporation | Screening small molecule drugs using neural cells differentiated from human embryonic stem cells |
WO2001088104A2 (en) | 2000-05-17 | 2001-11-22 | Geron Corporation | Neural progenitor cell populations |
US6576464B2 (en) | 2000-11-27 | 2003-06-10 | Geron Corporation | Methods for providing differentiated stem cells |
CA2453068C (en) | 2001-07-06 | 2018-02-20 | Geron Corporation | Mesenchymal cells and osteoblasts from human embryonic stem cell |
US7425448B2 (en) | 2001-07-12 | 2008-09-16 | Geron Corporation | Cardiomyocyte precursors from human embryonic stem cells |
CA2458362A1 (en) * | 2001-08-23 | 2003-03-06 | Reliance Life Sciences Pvt., Ltd. | Isolation of inner cell mass for the establishment of human embryonic stem cell (hesc) lines |
JP4148897B2 (ja) | 2001-10-31 | 2008-09-10 | 旭化成株式会社 | 胚性幹細胞培養用基材および培養方法 |
WO2003050250A2 (en) | 2001-12-07 | 2003-06-19 | Geron Corporation | Chondrocyte precursors derived from human embryonic stem cells |
US20030153082A1 (en) | 2001-12-07 | 2003-08-14 | The John P. Robarts Research Institute | Hematopoietic cells from human embryonic stem cells |
GB2415432B (en) | 2001-12-07 | 2006-09-06 | Geron Corp | Islet cells from human embryonic stem cells |
US20030113910A1 (en) | 2001-12-18 | 2003-06-19 | Mike Levanduski | Pluripotent stem cells derived without the use of embryos or fetal tissue |
US7285415B2 (en) | 2002-07-11 | 2007-10-23 | The Regents Of The University Of California | Oligodendrocytes derived from human embryonic stem cells for remyelination and treatment of spinal cord injury |
CN1483817A (zh) * | 2002-09-18 | 2004-03-24 | 李建远 | 一种克隆无动物成分的人胚胎干细胞的方法 |
ES2705683T3 (es) * | 2002-12-16 | 2019-03-26 | Technion Res & Dev Foundation | Medio de cultivo de células madre pluripotentes |
CN1424394A (zh) * | 2003-01-08 | 2003-06-18 | 中山大学附属第二医院 | 人胚胎干细胞系建立的方法 |
US20030224411A1 (en) | 2003-03-13 | 2003-12-04 | Stanton Lawrence W. | Genes that are up- or down-regulated during differentiation of human embryonic stem cells |
US7727762B2 (en) | 2003-10-03 | 2010-06-01 | Keiichi Fukuda | Method of inducing the differentiation of stem cells into myocardial cells |
CN100376670C (zh) * | 2003-12-10 | 2008-03-26 | 财团法人工业技术研究院 | 人类胚干细胞的培养系统及其培养方法 |
US20050233446A1 (en) * | 2003-12-31 | 2005-10-20 | Parsons Xuejun H | Defined media for stem cell culture |
GB2427873B (en) | 2004-03-19 | 2008-09-10 | Geron Corp | Method for making high purity cardiomyocyte preparations suitable for regenerative medicine |
DE102004043256B4 (de) * | 2004-09-07 | 2013-09-19 | Rheinische Friedrich-Wilhelms-Universität Bonn | Skalierbarer Prozess zur Kultivierung undifferenzierter Stammzellen in Suspension |
EP2410043A3 (en) | 2004-12-29 | 2013-01-23 | Hadasit Medical Research Services And Development Ltd. | Stem cells culture systems |
DE102005009751A1 (de) | 2005-03-03 | 2006-09-07 | Consortium für elektrochemische Industrie GmbH | Verfahren zur fermentativen Herstellung von S-Adenosyl-Methionin |
CN107189980B (zh) | 2005-06-22 | 2021-07-09 | 阿斯特利亚斯生物治疗股份公司 | 人胚胎干细胞的悬浮培养物 |
US8968994B2 (en) | 2006-07-06 | 2015-03-03 | Jeremy Micah Crook | Method for stem cell culture and cells derived therefrom |
PT2733203T (pt) | 2006-08-02 | 2019-01-23 | Technion Res & Dev Foundation | Métodos de expansão de células estminais embrionárias numa cultura de suspensão |
-
2006
- 2006-06-20 CN CN201710624640.3A patent/CN107189980B/zh active Active
- 2006-06-20 AU AU2006262369A patent/AU2006262369B2/en active Active
- 2006-06-20 KR KR1020157014388A patent/KR20150067394A/ko not_active Application Discontinuation
- 2006-06-20 US US11/917,993 patent/US9074181B2/en active Active
- 2006-06-20 EP EP06785185.7A patent/EP1910516B1/en active Active
- 2006-06-20 GB GB0800365A patent/GB2441488C/en active Active
- 2006-06-20 CN CN200680027460.7A patent/CN101233226B/zh active Active
- 2006-06-20 KR KR1020167026326A patent/KR20160116024A/ko not_active Application Discontinuation
- 2006-06-20 JP JP2008518312A patent/JP5560391B2/ja active Active
- 2006-06-20 KR KR1020087001755A patent/KR20080030039A/ko active Search and Examination
- 2006-06-20 CA CA2613369A patent/CA2613369C/en active Active
- 2006-06-20 KR KR1020137021804A patent/KR20130100221A/ko not_active Application Discontinuation
- 2006-06-20 WO PCT/US2006/023976 patent/WO2007002086A2/en active Application Filing
- 2006-06-20 EP EP19180013.5A patent/EP3599277A1/en active Pending
-
2007
- 2007-12-19 IL IL188264A patent/IL188264A/en active IP Right Grant
-
2008
- 2008-03-07 HK HK08102719.8A patent/HK1122836A1/xx unknown
-
2013
- 2013-02-26 JP JP2013035335A patent/JP5770213B2/ja active Active
-
2015
- 2015-07-06 US US14/791,479 patent/US20150307838A1/en not_active Abandoned
-
2018
- 2018-02-12 US US15/894,842 patent/US10676714B2/en active Active
-
2020
- 2020-04-29 US US16/862,559 patent/US20200399592A1/en active Pending
-
2024
- 2024-01-24 US US18/421,847 patent/US20240158741A1/en active Pending
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20240158741A1 (en) | Suspension culture of human embryonic stem cells | |
US7455983B2 (en) | Medium for growing human embryonic stem cells | |
AU2005271723B2 (en) | Medium for growing human embryonic stem cells | |
US7790456B2 (en) | Scalable process for cultivating undifferentiated stem cells in suspension | |
AU2002323593B2 (en) | Culture system for rapid expansion of human embryonic stem cells | |
AU2015249110B2 (en) | Suspension culture of human embryonic stem cells | |
Rothová et al. | Differentiation of mouse embryonic stem cells into ventral foregut precursors | |
AU2012203350B9 (en) | Suspension culture of human embryonic stem cells |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20090114 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110801 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110804 |
|
RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20110804 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110805 |
|
RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20110805 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20111018 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20111025 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120201 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20120202 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20121026 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20130226 |
|
A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20130412 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130628 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20130822 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20130829 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20140106 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20140228 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20140327 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20140328 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20140327 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20140521 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5560391 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
S531 | Written request for registration of change of domicile |
Free format text: JAPANESE INTERMEDIATE CODE: R313531 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |