CN105969878A - Application of long no-coding RNA ENST00000430247 - Google Patents
Application of long no-coding RNA ENST00000430247 Download PDFInfo
- Publication number
- CN105969878A CN105969878A CN201610430784.0A CN201610430784A CN105969878A CN 105969878 A CN105969878 A CN 105969878A CN 201610430784 A CN201610430784 A CN 201610430784A CN 105969878 A CN105969878 A CN 105969878A
- 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
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 long no-coding RNA (long no-coding RNA, LncRNA) ENST00000430247, that is, the long no-coding RNA is used for preparing a detection reagent for predicting differentiation of human umbilical cord sourced mesenchymal stem cells to osteoblast, and particularly, a kit for predicting differentiation of the human umbilical cord sourced mesenchymal stem cells to the osteoblast through a real-time fluorescent quantitative analysis method is prepared. It is proved through research that after the LncRNA ENST00000430247 is subjected to osteogenic induction culture treatment, high expression of ENST00000430247 is achieved. Therefore, profound clinical significance and popularization are achieved by applying expression of the ENST00000430247 to predicting of differentiation of the human umbilical cord sourced mesenchymal stem cells to the osteoblast.
Description
Technical field
The present invention relates to stem cell biology field, be specifically related to long-chain non-coding RNA ENST00000430247
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 immunoreation 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 ENST00000430247.LncRNA
ENST00000430247 (NONCODE ID:NONHSAT081598.2) is positioned No. 21 chromosomes, its expression energy conduct
Judge whether umbilical cord mesenchymal stem cells is induced to differentiate into osteoblastic foundation, therefore may be used for preparation prediction people's umbilical cord and come
The mescenchymal stem cell in source, to the preparation of osteoblast differentiation, further can provide a kind of cost performance high, it is easy to promoting should
The test kit of skeletonization efficiency rating.
The application of long-chain non-coding RNA ENST00000430247, does for preparing the mesenchyme in prediction people's umbilical cord source
Cell to the preparation of osteoblast differentiation, the transcript sequence such as SEQ NO of this long-chain non-coding RNA ENST00000430247:
Shown in 1.
It is non-that the mescenchymal stem cell in described prediction people's umbilical cord source includes detecting long-chain to the preparation of osteoblast differentiation
The real-time fluorescence quantitative PCR detectable of coding RNA ENST00000430247 expression.
Described real-time fluorescence quantitative PCR detectable includes the specific primer of real-time fluorescence quantitative PCR:
LncRNA ENST00000430247 specific forward primer:
5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 specific reverse primers:
5'-CTTTTCACCAGCCTCTCCAG-3'。
A kind of predict mescenchymal stem cell that people's umbilical cord originates to the test kit of osteoblast differentiation, this test kit includes:
LncRNA ENST00000430247 specific forward primer:
5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 specific reverse primers:
5'-CTTTTCACCAGCCTCTCCAG-3'。
This test 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 test 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 ENST00000430247 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 ENST00000430247, 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 ENST00000430247, find:
Inducing 14 days LncRNA ENST00000430247 to express is about 85 times not induced, and expresses and increase significantly after induction, other
Classical osteoblast molecule marker such as osteocalcin and osteopontin also increases, but multiple is not as LncRNA
The expression of ENST00000430247 is increased significantly.Accordingly, applicant proposes to utilize LncRNA ENST00000430247 to prepare
The mescenchymal stem cell in prediction people's umbilical cord source is to osteoblast differentiation reagent.
It is used for LncRNA ENST00000430247 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 ENST00000430247 reverse transcription as cDNA；(3) LncRNA is used
ENST00000430247 specific primer and internal reference primer carry out real-time fluorescence quantitative PCR amplification and obtain relative expression quantity, induction
The expression obvious difference do not induced, can as whether the prediction reagent of Osteoblast Differentiation.
The reagent utilizing the present invention can detect LncRNA ENST00000430247 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 ENST00000430247 osteogenic induction or do not induced
The differential expression of the mescenchymal stem cell in umbilical cord source；Although the expression induced 21 days will be less than induction 14 days, it may be possible to
Jointly regulate and control due to other genes or the factor to cause, but no matter the length of induction time will compare after still finding out induction
Expression before induction substantially increases；This lncRNA ENST00000430247 can be as umbilical cord mesenchymal stem cells to skeletonization
Cell differentiation mark；
Fig. 2 is skeletonization marker molecules OPN expression analysis；
QC1205con representative ordinary culture medium cultivates group；
QC1205diff representative Osteogenic Induction Medium cultivates group.
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 ENST00000430247
1. isopropanol 100ml
2.Trizol reagent 100ml
3. chloroform 50ml
4. 1 μM 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 ENST00000430247 specific primer 50 μ l
The specific primer of quantitative fluorescent PCR:
LncRNA ENST00000430247 forward primer: 5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 reverse primer: 5'-CTTTTCACCAGCCTCTCCAG-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 ENST00000430247 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:
Go 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 chloroform, 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 facies.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,
Add the RNA precipitate of water dissolution without RNase of 30～50 μ L, 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 ENST00000430247 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 ENST00000430247 specific primer and internal reference primer:
ENST00000430247 specific primer DNA sequence 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
ENST00000430247 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 ENST00000430247 forward primer: 5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 reverse primer: 5'-CTTTTCACCAGCCTCTCCAG-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 ENST00000430247 significantly raises, and difference has significance (p < 0.05).
More than research shows, lncRNA ENST00000430247 can divide to osteoblast as umbilical cord mesenchymal stem cells
Change mark.
Claims (6)
1. the application of long-chain non-coding RNA ENST00000430247, it is characterised in that for preparing prediction people's umbilical cord source
Mescenchymal stem cell is to the preparation of osteoblast differentiation, the transcript sequence of this long-chain non-coding RNA ENST00000430247
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 ENST00000430247 expression
Reagent.
Application the most according to claim 2, it is characterised in that described real-time fluorescence quantitative PCR detectable includes reality
Time quantitative fluorescent PCR specific primer:
LncRNA ENST00000430247 specific forward primer:
5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 specific reverse primers:
5'-CTTTTCACCAGCCTCTCCAG-3'。
4. predict that mescenchymal stem cell that people's umbilical cord originates is to osteoblast differentiation test kit for one kind, it is characterised in that this reagent
Box includes:
LncRNA ENST00000430247 specific forward primer:
5'-GCATTGGGATTACAGGGAGA-3'
LncRNA ENST00000430247 specific reverse primers:
5'-CTTTTCACCAGCCTCTCCAG-3'。
The most according to claim 4 prediction people's umbilical cord source mescenchymal stem cell to osteoblast differentiation test kit, its
Being characterised by, this test 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 osteoblast differentiation test kit,
It is characterized in that, this test 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 ENST00000430247 reverse transcription for cDNA agents useful for same, including reverse
Record buffer, 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 lncRNA ENST00000430247, real time fluorescent quantitative SYBR dyestuff, without enzyme water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610430784.0A CN105969878B (en) | 2016-06-17 | 2016-06-17 | The application of long-chain non-coding RNA ENST00000430247 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610430784.0A CN105969878B (en) | 2016-06-17 | 2016-06-17 | The application of long-chain non-coding RNA ENST00000430247 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105969878A true CN105969878A (en) | 2016-09-28 |
| CN105969878B CN105969878B (en) | 2019-03-05 |
Family
ID=57022703
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610430784.0A Expired - Fee Related CN105969878B (en) | 2016-06-17 | 2016-06-17 | The application of long-chain non-coding RNA ENST00000430247 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105969878B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108004311A (en) * | 2017-12-20 | 2018-05-08 | 中国人民解放军第四军医大学 | The application of long-chain non-coding RNA NONMMUT002009 and its overexpression plasmid in diagnose and treat disease of skeletal system |
| CN112391386A (en) * | 2020-11-24 | 2021-02-23 | 梁树卷 | Mesenchymal stem cell migration promoter |
-
2016
- 2016-06-17 CN CN201610430784.0A patent/CN105969878B/en not_active Expired - Fee Related
Non-Patent Citations (6)
| Title |
|---|
| ANONYMOUS: "ENST00000430247", 《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》 * |
| RAMIRO GARZON ET AL: "Expression and prognostic impact of lncRNAs in acute myeloid leukemia", 《PNAS》 * |
| 孙翔等: "人骨髓间充质干细胞成骨分化相关的长链非编码RNA表达谱特征", 《广东医学》 * |
| 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 (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108004311A (en) * | 2017-12-20 | 2018-05-08 | 中国人民解放军第四军医大学 | The application of long-chain non-coding RNA NONMMUT002009 and its overexpression plasmid in diagnose and treat disease of skeletal system |
| CN108004311B (en) * | 2017-12-20 | 2021-08-03 | 中国人民解放军第四军医大学 | Long-chain non-coding RNA NONMMUT002009 and application of overexpression plasmid thereof in diagnosis and treatment of bone system diseases |
| CN112391386A (en) * | 2020-11-24 | 2021-02-23 | 梁树卷 | Mesenchymal stem cell migration promoter |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105969878B (en) | 2019-03-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Haas et al. | Advances in decoding axolotl limb regeneration | |
| Czarkwiani et al. | Expression of skeletogenic genes during arm regeneration in the brittle star Amphiura filiformis | |
| Mandáková et al. | Painting of Arabidopsis chromosomes with chromosome‐specific BAC clones | |
| CN105886509B (en) | The application of long-chain non-coding RNA ENST00000429456 | |
| JP2018075030A (en) | Measurement of telomere length in formalin fixed paraffin embedded (ffpe) sample by quantitive pcr | |
| Li et al. | Molecular characterization of heterogeneous mesenchymal stem cells with single-cell transcriptomes | |
| Dong et al. | Comparison of long noncoding RNA and mRNA expression profiles in mesenchymal stem cells derived from human periodontal ligament and bone marrow | |
| Krešić et al. | Canine adipose derived mesenchymal stem cells transcriptome composition alterations: a step towards standardizing therapeutic | |
| CN105969878B (en) | The application of long-chain non-coding RNA ENST00000430247 | |
| Zhang et al. | Gene expression of single human mesenchymal stem cell in response to fluid shear | |
| CN105925706B (en) | The application of long-chain non-coding RNA LOC100506530 | |
| CN105671175A (en) | Detection method for cattle SPAG17 gene insertion/deletion and application thereof | |
| CN105925707B (en) | The application of long-chain non-coding RNA ENST00000520431 | |
| CN105886510B (en) | The application of long-chain non-coding RNA ENST00000529841 | |
| CN104109669B (en) | Genetic markers and application of the pig AMPD1 gene promoter regions SNP as hog on hook character | |
| CN1978664A (en) | Method for identifying goat early embryo sex via PCR | |
| WO2019240073A1 (en) | Quantitative pcr probe | |
| CN107287227A (en) | A kind of goat miR 27a pointed decorations systems and its application | |
| CN105950754B (en) | The application of long-chain non-coding RNA ENST00000511346 | |
| CN103740842B (en) | Highly sensitive tiny RNA quantitative detecting method and test kit | |
| Hoikkala et al. | Cooperation between CRISPR-Cas types enables adaptation in an RNA-targeting system | |
| Lakshmipathy et al. | Micro RNA profiling: an easy and rapid method to screen and characterize stem cell populations | |
| Perán et al. | Cellular extracts from post-mortem human cardiac tissue direct cardiomyogenic differentiation of human adipose tissue-derived stem cells | |
| GB2550139A (en) | Functional prediction of MSCs | |
| CN107312775B (en) | Application of hsa _ circRNA _103096 in diagnosis, treatment and prognosis of liver cancer |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190305 Termination date: 20200617 |