CN105925707A - Application of long non-coding RNAENST00000520431 - Google Patents

Application of long non-coding RNAENST00000520431 Download PDF

Info

Publication number
CN105925707A
CN105925707A CN201610435777.XA CN201610435777A CN105925707A CN 105925707 A CN105925707 A CN 105925707A CN 201610435777 A CN201610435777 A CN 201610435777A CN 105925707 A CN105925707 A CN 105925707A
Authority
CN
China
Prior art keywords
lncrna
primer
umbilical cord
stem cell
mescenchymal stem
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.)
Granted
Application number
CN201610435777.XA
Other languages
Chinese (zh)
Other versions
CN105925707B (en
Inventor
帅词俊
彭淑平
高丹
何世微
钟雁城
曾朝阳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Central South University
Original Assignee
Central South University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Central South University filed Critical Central South University
Priority to CN201610435777.XA priority Critical patent/CN105925707B/en
Publication of CN105925707A publication Critical patent/CN105925707A/en
Application granted granted Critical
Publication of CN105925707B publication Critical patent/CN105925707B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6851—Quantitative amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses application of LncRNA (long non-coding RNA) ENST00000520431 to preparation of a detection reagent used for predicating differentiation of human umbilical cord derived mesenchymal stem cells to osteoblasts and preparation of a kit for predicating differentiation of human umbilical cord derived mesenchymal stem cells to osteoblasts according to a real-time fluorescence quantification method. According to researches, after osteogenic induction culture treatment of LncRNA ENST00000520431, ENST00000520431 expression is remarkably reduced. Therefore, application of ENST00000520431 expression to predication of differentiation of mesenchymal stem cells to osteoblasts has profound clinical significance and popularization.

Description

The application of long-chain non-coding RNA ENST00000520431
Technical field
The present invention relates to stem cell biology field, be specifically related to long-chain non-coding RNA ENST00000520431 and exist The application in osteoblast differentiation preparation of the mescenchymal stem cell in preparation prediction people's umbilical cord source.
Background technology
Mescenchymal stem cell (Mesenchymal stem cells, MSCs) is that one has the of self-replication capacity and multidirectional The adult stem cell of differentiation potential, belongs to non-terminally differentiated cells, in vitro under specific inductive condition, can be divided into fat, The Various Tissues cells such as cartilage, bone, muscle, tendon, nerve, liver, cardiac muscle, beta Cell of islet and endothelium, continuous passage is cultivated and cold Still there is after freezing preservation multi-lineage potential, be a kind of preferably Seeding Cells in Bone Tissue Engineering.Current most widely used bone The mescenchymal stem cell in the source such as marrow, fat, umbilical cord and Cord blood, they are respectively provided with multi-lineage potential；And be easily obtained And amplification, still there is after continuous passage cultivation and freezen protective multi-lineage potential, can be that reparation and the regeneration of histoorgan carries For required a large amount of cells；Immunogenicity is low simultaneously, the most autologous or allogenic mescenchymal stem cell, the most not The immune response of host can be caused, also there is the immunoloregulation function of uniqueness.
What long-chain non-coding RNA (Long non-coding RNA, lncRNAs) was a class more than 200bp does not encodes egg White RNA.Along with the universal of high throughput sequencing technologies and the understanding to LncRNAs, it has been found that LncRNAs is in gene expression Regulation and control play very important effect.Same during mescenchymal stem cell directed differentiation, LncRNA is also by regulation and control The expression of related gene affects the direction of differentiation.
Summary of the invention
It is an object of the invention to provide the application of a kind of LncRNA ENST00000520431.LncRNA ENST00000520431 (NONCODE ID:NONHSAT126033.2) is positioned No. 8 chromosomes, and its expression can be as sentencing Whether disconnected umbilical cord mesenchymal stem cells is induced to differentiate into osteoblastic foundation, therefore may be used for preparation prediction people's umbilical cord source Mescenchymal stem cell to the preparation of osteoblast differentiation, a kind of cost performance further can be provided high, it is easy to popularization and application The kit of skeletonization efficiency rating.
The application of long-chain non-coding RNA ENST00000520431, does for preparing the mesenchyma in prediction people's umbilical cord source Cell is to osteoblast differentiation preparation, the transcript sequence such as SEQ NO:1 of this long-chain non-coding RNA ENST00000520431 Shown in.
The mescenchymal stem cell in described prediction people's umbilical cord source includes detecting the non-volume of long-chain to osteoblast differentiation preparation The real-time fluorescence quantitative PCR detection reagent of code RNA ENST00000520431 expression.
Described real-time fluorescence quantitative PCR detection reagent includes the specific primer of real-time fluorescence quantitative PCR:
LncRNA ENST00000520431 specific forward primer:
5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 specific reverse primers:
5'-TCTTGTCCTCGCCTTGTCTT-3'。
A kind of predict mescenchymal stem cell that people's umbilical cord originates to the kit of osteoblast differentiation, this kit includes:
LncRNA ENST00000520431 specific forward primer:
5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 specific reverse primers:
5'-TCTTGTCCTCGCCTTGTCTT-3'。
This kit also includes:
The specific primer of internal reference β-actin:
Forward primer 5'-CCTATCGAGCATGGAGTGGT-3'
Reverse primer 5'-CTGAGGCATAGAGGGACAGC-3'
The specific primer of internal reference α-Tubulin:
Forward primer 5'-CCTGATGTATGCCAAACGTG-3'
Reverse primer 5'-TCCCTGTAAAAGCAGCACCT-3'.
This kit complete set reagent includes:
(1) from the mescenchymal stem cell extracted total RNA agents useful for same of induction, including Trizol reagent, chloroform, isopropyl Alcohol, without enzyme water；(2) with total serum IgE for template by lncRNA ENST00000520431 reverse transcription for cDNA agents useful for same, including inverse Transcription buffer, dNTP, RNase inhibitor, MMLV reverse transcriptase and random primer；(3) by used by cDNA real-time quantitative PCR Reagent, including the specific primer of lncRNA ENST00000520431, real time fluorescent quantitative SYBR dyestuff, without enzyme water.
The present invention, by the mescenchymal stem cell in Osteogenic Induction Medium induction people's umbilical cord source, after processing 1,2,3 weeks, carries Take cell total rna, carry out real-time fluorescence quantitative PCR after reverse transcription and analyze the expression of LncRNA ENST00000520431, find: The LncRNA ENST00000520431 not induced is about 20 times that induction is expressed for 14 days, expresses and reduce significantly after induction, its Gegenbaur's cell molecule marker such as osteocalcin and osteopontin of its classics then increases.Accordingly, applicant proposes profit The mescenchymal stem cell in prediction people's umbilical cord source is prepared to osteoblast differentiation reagent with LncRNA ENST00000520431.
It is used for LncRNA ENST00000520431 predicting that the mescenchymal stem cell that people's umbilical cord is originated is the thinnest to skeletonization The method of born of the same parents' differentiation: the mescenchymal stem cell in people's umbilical cord source that (1) is collected after osteogenic induction or do not induced, extracted total RNA； (2) with total serum IgE for template by LncRNA ENST00000520431 reverse transcription as cDNA；(3) LncRNA is used ENST00000520431 specific primer and internal reference primer carry out real-time fluorescence quantitative PCR amplification and obtain relative expression quantity, induction The expression difference do not induced is obvious, can as whether the prediction reagent of Osteoblast Differentiation.
The reagent utilizing the present invention can detect LncRNA ENST00000520431 people's umbilical cord after osteogenic induction The expression of the mescenchymal stem cell in source thus judge whether Osteoblast Differentiation, carry for human umbilical cord mesenchymal stem cells Osteoblast Differentiation Supply the molecular marker of lncRNA, there is far-reaching practice significance and generalization.
Illustrate to further illustrate the present invention with detailed description of the invention below in conjunction with accompanying drawing, and the unrestricted present invention.
Accompanying drawing explanation
Fig. 1 is the people that real-time fluorescence quantitative PCR is analyzed after LncRNA ENST00000520431 osteogenic induction or do not induced The differential expression of the mescenchymal stem cell in umbilical cord source；
QC1205con representative ordinary culture medium cultivates group；
QC1205diff representative Osteogenic Induction Medium cultivates group.
Fig. 2 is skeletonization marker molecules OCN expression analysis.
Detailed description of the invention
LncRNA in the human umbilical cord mesenchymal stem cells that embodiment 1 Osteogenic Induction Medium is induced 1,2,3 weeks and do not induced The detection kit of ENST00000520431
1. isopropanol 100ml
2.Trizol reagent 100ml
3. chloroform 50ml
4.1 μMs of random reverse transcriptase primer 50 μ l
5. without enzyme water 2ml
6.10mM dNTP 100μl
7.200U/ μ l RNA reverse transcriptase 50 μ l
8.5 × RT Buffer 1ml
9.40U/ μ l RNA inhibitor 500 μ l
10.Premix Ex Taq 50μl
11.10 μMs of lncRNA ENST00000520431 specific primer 50 μ l
The specific primer of quantitative fluorescent PCR:
LncRNA ENST00000520431 forward primer: 5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 reverse primer: 5'-TCTTGTCCTCGCCTTGTCTT-3'
12.10 μMs of internal reference comparison each 50 μ l of primer
The specific primer of internal reference β-actin:
5'-CCTATCGAGCATGGAGTGGT-3'
5'-CTGAGGCATAGAGGGACAGC-3'
The specific primer of internal reference α-Tubulin:
5'-CCTGATGTATGCCAAACGTG-3'
5'-TCCCTGTAAAAGCAGCACCT-3'
The detection of LncRNA ENST00000520431 after embodiment 2 human umbilical cord mesenchymal stem cells osteogenic induction
(1) mescenchymal stem cell in people's umbilical cord source that osteogenic induction 1,2,3 Zhou Houhe does not induces is collected, extracted total RNA: Remove culture medium in the culture plate after most induction, add 1ml Trizol, at room temperature horizontal positioned 5min, make lysate uniformly divide It is distributed in cell surface, then makes cell detachment with liquid-transfering gun piping and druming cell；Cell pyrolysis liquid is transferred in centrifuge tube, adds 200 μ l chloroforms, cover tightly centrifuge tube lid, up and down vibration 15 seconds, and room temperature stands 3 minutes, 12,000g, 4 DEG C centrifugal 15 points Clock.Taking out centrifuge tube, sample is divided into three layers: the supernatant aqueous phase of light color, middle white layer and peach lower floor organic phase.Little The heart is drawn light color supernatant water and is moved to another centrifuge tube mutually, adds isopyknic isopropanol, mixes gently, and room temperature stands 10 minutes, Then 12,000g, 4 DEG C centrifugal 10 minutes, RNA precipitate seen from the bottom of pipe.Carefully remove supernatant, slowly add along tube wall 1mL75% ethanol, mixes gently.12,000g, 4 DEG C are centrifuged 10 minutes, carefully exhaust supernatant.Drying at room temperature precipitates 2～5 minutes, The water without RNase adding 30～50 μ L dissolves RNA precipitate, the concentration of spectrophotometer detection RNA and quality, OD260/280 ratio It is worth between 1.8-2.0 ,-70 DEG C of preservations.
(2) with total serum IgE for template by LncRNA ENST00000520431 reverse transcription as cDNA；
Oligo(dT)18primer(100μM) 1μl
Total RNA 1μg
The Total of water without enzyme 12 μ l
Reverse transcription first step condition: 62 DEG C 5 minutes
5 × RT Buffer 4 μ l
dNTP(10mM)2μl
RNase inhibitor (40U/ μ l) 1 μ l
MMLV reverse transcriptase (200U/ μ l) 1 μ l
First step product 12 μ l
Total 20μl
Reverse transcription second step program, 42 DEG C 60 minutes, 72 DEG C 5 minutes.
(3) real-time fluorescence quantitative PCR is carried out with LncRNA ENST00000520431 specific primer and internal reference primer: ENST00000520431 specific primer DNA sequence dna is synthesized by Invitrogen company.
First reverse transcription product is diluted 5 times, mixing, 20 μ l reaction systems are as follows:
SYBR Premix 2×10μl
ENST00000520431 or internal reference primer 1 μ l
CDNA product 0.5 μ l
The Total of water without enzyme 20 μ l
Real-time fluorescence quantitative PCR react 95 DEG C 30 seconds, 40 circulation 95 DEG C 5 seconds, 65 DEG C 30 seconds, 72 DEG C 30 seconds, 65～ 95 DEG C, within every 0.05 second, increase by 0.5 DEG C.
LncRNA ENST00000520431 forward primer: 5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 reverse primer: 5'-TCTTGTCCTCGCCTTGTCTT-3'
The specific primer of internal reference β-actin:
Forward primer 5'-CCTATCGAGCATGGAGTGGT-3'
Reverse primer 5'-CTGAGGCATAGAGGGACAGC-3'
The specific primer of internal reference α-Tubulin:
Forward primer 5'-CCTGATGTATGCCAAACGTG-3'
Reverse primer 5'-TCCCTGTAAAAGCAGCACCT-3'
(4) mensuration of osteogenic induction: this experimental data uses the analysis method of relative quantification, β-actin and α-Tubulin As reference gene, data separate GraphPad Prism is analyzed.Success inducing umbilical cord mesenchymal stem is thin to skeletonization After born of the same parents' differentiation, lncRNA ENST00000520431 significantly lowers, and difference has conspicuousness (p < 0.05).
More than research shows, lncRNA ENST00000520431 can divide to Gegenbaur's cell as umbilical cord mesenchymal stem cells Change mark.

Claims (6)

1. the application of long-chain non-coding RNA ENST00000520431, it is characterised in that for preparing prediction people's umbilical cord source Mescenchymal stem cell is to the preparation of osteoblast differentiation, and the transcript sequence of this long-chain non-coding RNA ENST00000520431 is such as Shown in SEQ NO:1.
Application the most according to claim 1, it is characterised in that the mescenchymal stem cell in prediction people's umbilical cord source is thin to skeletonization The preparation of born of the same parents' differentiation includes the real-time fluorescence quantitative PCR detection detecting long-chain non-coding RNA ENST00000520431 expression Reagent.
Application the most according to claim 2, it is characterised in that described real-time fluorescence quantitative PCR detection reagent includes reality Time quantitative fluorescent PCR specific primer:
LncRNA ENST00000520431 specific forward primer:
5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 specific reverse primers:
5'-TCTTGTCCTCGCCTTGTCTT-3'。
4. predict that mescenchymal stem cell that people's umbilical cord originates is to the kit of osteoblast differentiation for one kind, it is characterised in that this examination Agent box includes:
LncRNA ENST00000520431 specific forward primer:
5'-GGCACATGGAGAATTAAATG-3'
LncRNA ENST00000520431 specific reverse primers:
5'-TCTTGTCCTCGCCTTGTCTT-3'。
The most according to claim 4 prediction people's umbilical cord source mescenchymal stem cell to osteoblast differentiation kit, its Being characterised by, this kit includes:
The specific primer of internal reference β-actin:
Forward primer 5'-CCTATCGAGCATGGAGTGGT-3'
Reverse primer 5'-CTGAGGCATAGAGGGACAGC-3'
The specific primer of internal reference α-Tubulin:
Forward primer 5'-CCTGATGTATGCCAAACGTG-3'
Reverse primer 5'-TCCCTGTAAAAGCAGCACCT-3'.
6. according to the mescenchymal stem cell predicting people's umbilical cord source described in claim 4 or 5 to the reagent of osteoblast differentiation Box, it is characterised in that this kit includes:
(1) from the mescenchymal stem cell extracted total RNA agents useful for same of induction, including Trizol reagent, chloroform, isopropanol, Water without enzyme；(2) with total serum IgE for template by lncRNA ENST00000520431 reverse transcription for cDNA agents useful for same, including reverse Record buffer solution, dNTP, RNase inhibitor, MMLV reverse transcriptase and random primer；(3) will try used by cDNA real-time quantitative PCR Agent, including the specific primer of lncRNAENST00000520431, real time fluorescent quantitative SYBR dyestuff, without enzyme water.
CN201610435777.XA 2016-06-17 2016-06-17 The application of long-chain non-coding RNA ENST00000520431 Expired - Fee Related CN105925707B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610435777.XA CN105925707B (en) 2016-06-17 2016-06-17 The application of long-chain non-coding RNA ENST00000520431

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610435777.XA CN105925707B (en) 2016-06-17 2016-06-17 The application of long-chain non-coding RNA ENST00000520431

Publications (2)

Publication Number Publication Date
CN105925707A true CN105925707A (en) 2016-09-07
CN105925707B CN105925707B (en) 2019-02-22

Family

ID=56830611

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610435777.XA Expired - Fee Related CN105925707B (en) 2016-06-17 2016-06-17 The application of long-chain non-coding RNA ENST00000520431

Country Status (1)

Country Link
CN (1) CN105925707B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109136228A (en) * 2018-10-10 2019-01-04 新乡医学院 Application of the long-chain non-coding RNA-NKILA in bone tissue injury repair

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "LINC01605:16", 《互联网文章》 *
MAN NIU ET AL: "MiR-29c regulates the expression of miR-34c and miR-449a by targeting DNA methyltransferase 3a and 3b in nasopharyngeal carcinoma", 《BMC CANCER》 *
张红艳等: "利用共培养法诱导人胎盘间充质干细胞向成骨细胞分化", 《中华骨质疏松和骨矿盐疾病杂志》 *
邵帅等: "体外诱导骨髓与脐血间充质干细胞向成骨细胞分化及其成骨活性的研究", 《中国组织工程研究》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109136228A (en) * 2018-10-10 2019-01-04 新乡医学院 Application of the long-chain non-coding RNA-NKILA in bone tissue injury repair
CN109136228B (en) * 2018-10-10 2021-11-12 新乡医学院 Application of long-chain non-coding RNA-NKILA in bone tissue injury repair

Also Published As

Publication number Publication date
CN105925707B (en) 2019-02-22

Similar Documents

Publication Publication Date Title
Haas et al. Advances in decoding axolotl limb regeneration
Tesson et al. Ice nucleation activity and aeolian dispersal success in airborne and aquatic microalgae
Lavania et al. Detection of viable Mycobacterium leprae in soil samples: insights into possible sources of transmission of leprosy
CN105886509A (en) Application of long no-coding RNA ENST00000429456
Trifonov et al. Chromosomal evolution in Gekkonidae. I. Chromosome painting between Gekko and Hemidactylus species reveals phylogenetic relationships within the group
Kang et al. Genetic comparison of stemness of human umbilical cord and dental pulp
Dong et al. Comparison of long noncoding RNA and mRNA expression profiles in mesenchymal stem cells derived from human periodontal ligament and bone marrow
Sun et al. 5‐Azacytidine‐Induced Cardiomyocyte Differentiation of Very Small Embryonic‐Like Stem Cells
CN105969878B (en) The application of long-chain non-coding RNA ENST00000430247
CN105925706B (en) The application of long-chain non-coding RNA LOC100506530
CN105925707B (en) The application of long-chain non-coding RNA ENST00000520431
Lo et al. Arabiibacter massiliensis gen. nov. sp. nov., new anaerobic bacterium isolated from the human gut
CN105886510A (en) Application of long no-coding RNA ENST00000529841
CN104109669B (en) Genetic markers and application of the pig AMPD1 gene promoter regions SNP as hog on hook character
CN105950754B (en) The application of long-chain non-coding RNA ENST00000511346
Kostyuk et al. Fragments of cell-free DNA increase transcription in human mesenchymal stem cells, activate TLR-dependent signal pathway, and suppress apoptosis
Hoikkala et al. Cooperation between CRISPR-Cas types enables adaptation in an RNA-targeting system
Kagami et al. Fundamental technological developments required for increased availability of tissue engineering
Perán et al. Cellular extracts from post-mortem human cardiac tissue direct cardiomyogenic differentiation of human adipose tissue-derived stem cells
Kalfeist et al. Simultaneous isolation of CD45 tumor-infiltrating lymphocytes, tumor cells, and associated fibroblasts from murine breast tumor model by MACS
CN107312775B (en) Application of hsa _ circRNA _103096 in diagnosis, treatment and prognosis of liver cancer
Singh et al. In vitro culture and characterization of testis-derived cells from Clarias magur (Hamilton, 1822)
CN115807065B (en) Method for detecting drug-resistant predatory mite population by heat shock protein Hsp70 gene
Yakushiji-Kaminatsui et al. Genetic, genomic, and imaging approaches to dissect the role of Polycomb group epigenetic regulators in mice
Afouda Stem-cell-like embryonic explants to study cardiac development

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20190222

Termination date: 20190617

CF01 Termination of patent right due to non-payment of annual fee