CN106244520A - The preparation method of metanephros mesenchymal cell - Google Patents
The preparation method of metanephros mesenchymal cell Download PDFInfo
- Publication number
- CN106244520A CN106244520A CN201610630153.3A CN201610630153A CN106244520A CN 106244520 A CN106244520 A CN 106244520A CN 201610630153 A CN201610630153 A CN 201610630153A CN 106244520 A CN106244520 A CN 106244520A
- Authority
- CN
- China
- Prior art keywords
- foxd1
- cre
- embryo
- dtr
- flox
- 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
- 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
-
- 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
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Developmental Biology & Embryology (AREA)
- Reproductive Health (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Gynecology & Obstetrics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides the preparation method of a kind of metanephros mesenchymal cell, by by Foxd1creTransgenic mice and DTRfloxTransgenic mice carries out copulation, separates embryo Ren Mus dirty, culturing embryo nephrocyte, extracts highly purified Foxd1 by adding diphtheria toxin, diphtherotoxin+Metanephros mesenchymal cell.Compared with the method separating specific cells phenotype metanephros mesenchymal cell reported at present, this method is simple to operate, the Foxd1 extracted+Metanephros mesenchymal cell composition is single, and purity is high；Foxd1+Metanephros mesenchymal cell extracted amount is controlled, can realize Foxd1+Prepared by the large-scale production of metanephros mesenchymal cell.
Description
Technical field
The present invention relates to genetic engineering and biology field, specifically, relate to the preparation of metanephros mesenchymal cell
Method.
Background technology
Mammal kidney is grown with the interaction of metanephros mesenchyme by metanephros ureteric bud, and the former develops into
For ureter and collecting tubule, the latter is divided into two groups of cells, i.e. fills between hat shape mesenchymal cell (mark is Six2) and interstitial
Cell plastid (mark is Foxd1).In kidney development/embryonic kidney cells correlational study, it is sometimes desirable to by metanephros mesenchymal cell,
The even metanephros mesenchymal cell of specific cells phenotype is separated, and research previously only takes out embryo in required specified number of days
Kidney, cut off, culture of isolated embryonic kidney cells, if desired for the rear nephrocyte of specific cells phenotype (with Foxd1+As a example by cell), will divide
From metanephros mesenchymal cell carry out Foxd1 fluorescence staining, then by flow cytometry screening and separating Foxd1+Cell.
The method not only complex steps separating specific cells phenotype metanephros mesenchymal cell reported at present, and exist thin
The problems such as born of the same parents' composition is uncertain, cell yield is low.
Summary of the invention
It is an object of the invention to provide the preparation method of a kind of metanephros mesenchymal cell.
In order to realize the object of the invention, the present invention is by by Foxd1creTransgenic mice and DTRfloxTransgenic mice enters
Row copulation, separates embryo Ren Mus dirty, culturing embryo nephrocyte, extracts the highly purified mark Foxd1 positive by adding diphtheria toxin, diphtherotoxin
(Foxd1+) metanephros mesenchymal cell.
The preparation method of the metanephros mesenchymal cell that the present invention provides, by Foxd1creTransgenic mice and DTR (diphtheria poison
Element receptor)floxTransgenic mice carries out copulation, obtain female Mus become pregnant start after the embryo in 13.5-15.5 days (preferably,
It is designated as E0.5 days being found to have vaginal suppository, female Mus is fed to the E13.5-15.5 days), separate embryonic kidney, embryonic kidney is shredded and is followed by
Planting and cultivate in stem cell special culture media, identify the genotype of each embryo simultaneously, filtering out genotype is
Foxd1-cre；The double transgenic type embryonic kidney of DTR-flox, cultivates addition final concentration 100-in 60-72 hour backward culture medium
The diphtheria toxin, diphtherotoxin of 150ng/ml (preferably 100ng/ml), every 48 hours with containing 100-150ng/ml (preferably 100ng/ml) diphtheria poison
The stem cell special culture media of element changes liquid 1 time, until cultivating 5-7 days, the survivaling cell collected is Foxd1+Fill between metanephros
Cell plastid.
Wherein, Foxd1creTransgenic mice is purchased from U.S.'s Jackson laboratory (Stock No:012463).
DTRfloxTransgenic mice by Yale University Lloyd professor Cantley give (Guo J K, Shi H,
Koraishy F,et al.The Terminator mouse is a diphtheria toxin–receptor knock-in
mouse strain for rapid and efficient enrichment of desired cell lineages[J]
.Kidney international,2013,84(5):1041-1046.)。
The stem cell special culture media related in the present invention is that C57BL/6 Marrow Mesenchymal Stem Cells is cultivated completely
Base, is produced by match industry (Guangzhou) bio tech ltd, goods number MUBMX-90011.
Aforesaid method, embryonic kidney is inoculated in after shredding in stem cell special culture media, in 37 DEG C, 5%CO2Under the conditions of cultivate
2-3 days.
Aforesaid method, identifies the genotype of each embryo, and screening-gene type is Foxd1-cre；DTR-flox's
The concrete grammar of double transgenic type embryonic kidney is as follows, comprises the following steps:
The qualification of S1, Foxd1-cre transgenic mice
S11, embryo Mus rat-tail extracting genome DNA；
S12, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, and 2
× EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s,
65 DEG C of 30s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S13, result detect: prepare 1.5% agarose gel, and every hole adds PCR product 5 μ l, under 140V voltage
Electrophoresis 20min, observes electricity kank fruit in gel imaging system, if there is the amplified band of about 450bp size, is then accredited as
Foxd1-cre transgenic mice；
Wherein, in S12, the primer sequence is as follows:
Forward primer F1 5 '-TCTGGTCCAAGAATCCGAAG-3 ', downstream primer R1 5 '-
GGGAGGATTGGGAAGACAAT-3’；
S2、Foxd1-cre；The qualification of DTR-flox double transgenic type mice
S21, it is accredited as the embryo of Foxd1-cre transgenic mice as material with above-mentioned, extracts embryo Mus rat-tail genome
DNA；
S22, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, and 2
× EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s,
50 DEG C of 45s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S23, preparing 1.5% agarose gel, every hole adds PCR product 5 μ l, electrophoresis 20min under 140V voltage,
In gel imaging system, observe electricity kank fruit, if there is the amplified band of about 650bp size, be then accredited as Foxd1-cre；
DTR-flox double transgenic type mice；
Wherein, in S22, the primer sequence is as follows:
Forward primer F2 5 '-ACCATGAAGCTGCTGCCGTC-3 ', downstream primer R2 5 '-
TCAGTGGGAATTAGTCATGC-3’；
S3, it is accredited as Foxd1-cre from above-mentioned；The embryo of DTR-flox double transgenic type mice separates embryonic kidney, is
Genotype is Foxd1-cre；The double transgenic type embryonic kidney of DTR-flox.
The present invention is with Foxd1creTransgenic mice and DTRfloxBased on transgenic mice, obtain embryo Mus by copulation,
Foxd-cre；In DTR-flox double transgenic embryo Mus body, the cell not expressing Foxd1 will express DTR, this kind of cells contacting diphtheria
By death after toxin, and the cell expressing Foxd1 does not express DTR, still survives (Fig. 1) after contact diphtheria toxin, diphtherotoxin.Embryonic kidney Foxd1+
Metanephros mesenchymal cell is had an effect at Cre with loxP, makes the DTRpA genetic fragment (diptheria toxin receptor) being positioned between loxP sink
Silent, thus do not express DTR, therefore reactionless to diphtheria toxin, diphtherotoxin；Rather than Foxd1+Other embryonic kidney cells, due to loxP-
DTRpA-loxP continues to express DTR, and after adding diphtheria toxin, diphtherotoxin, cell is by death.This principle is utilized to extract highly purified Foxd1+
Metanephros mesenchymal cell.Carry out extracting the metanephros separated without culture medium adds diphtheria toxin, diphtherotoxin with Real-time PCR detection
The Foxd1 that mesenchymal cell and employing said method extract+The expression of Foxd1 and Six2 of metanephros mesenchymal cell,
Result is as in figure 2 it is shown, Foxd1+Metanephros mesenchymal cell high expressed Foxd1, and express Six2 hardly, thus may determine that logical
Cross the inventive method and successfully extract highly purified Foxd1+Metanephros mesenchymal cell.
The invention have the advantages that
(1) compared with the method separating specific cells phenotype metanephros mesenchymal cell reported at present, this method operates
Simply, the Foxd1 extracted+Metanephros mesenchymal cell composition is single, and purity is high；
(2) Foxd1+Metanephros mesenchymal cell extracted amount is controlled, can realize Foxd1+Metanephros mesenchymal cell extensive
Produce preparation.
Accompanying drawing explanation
Fig. 1 is Foxd1 of the present invention+The preparation principle of metanephros mesenchymal cell.
Fig. 2 is Foxd1 in the embodiment of the present invention 1+Metanephros mesenchymal cell is with metanephros mesenchymal cell Foxd1's and Six2
Expression compares.
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.If not specializing, embodiment
In the conventional means that is well known to those skilled in the art of technological means used, raw materials used be commercial goods.
Embodiment 1 Foxd1+The preparation of metanephros mesenchymal cell
First by Foxd1creTransgenic mice and DTRfloxTransgenic mice carries out copulation, and every day, twice observation sooner or later was female
Whether Mus becomes pregnant (whether having vaginal suppository), is designated as E0.5 days as being found to have vaginal suppository, is taken out by female Mus and individually feeds until the
E13.5-15.5 days, after neck that female Mus is broken execution, sterilization, take out uterus immediately, separate embryo, with micro-under direct-view microscope
Operating theater instruments separation embryonic kidney, during the embryonic kidney of homeomorphism Mus is not respectively put into the EP pipe filling normal saline, the rat-tail of its corresponding embryo Mus
The EP pipe being also placed in numbering corresponding treats that genotype identification is standby.Embryonic kidney is shredded and is inoculated in stem cell special culture media, in 37
DEG C, 5%CO2Under the conditions of cultivate, the genotype of each embryo Mus is identified simultaneously, filtering out genotype is Foxd1-cre；DTR-
The embryonic kidney of flox double transgenic type.Concrete grammar is as follows:
The qualification of S1, Foxd1-cre transgenic mice
S11, embryo Mus rat-tail extracting genome DNA；
S12, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, and 2
× EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s,
65 DEG C of 30s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S13, result detect: prepare 1.5% agarose gel, and every hole adds PCR product 5 μ l, under 140V voltage
Electrophoresis 20min, observes electricity kank fruit in gel imaging system, if there is the amplified band of about 450bp size, is then accredited as
Foxd1-cre transgenic mice；
Wherein, in S12, the primer sequence is as follows:
Forward primer F1 5 '-TCTGGTCCAAGAATCCGAAG-3 ', downstream primer R1 5 '-
GGGAGGATTGGGAAGACAAT-3’；
S2、Foxd1-cre；The qualification of DTR-flox double transgenic type mice
S21, it is accredited as the embryo of Foxd1-cre transgenic mice as material with above-mentioned, extracts embryo Mus rat-tail genome
DNA；
S22, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, and 2
× EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s,
50 DEG C of 45s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S23, preparing 1.5% agarose gel, every hole adds PCR product 5 μ l, electrophoresis 20min under 140V voltage,
In gel imaging system, observe electricity kank fruit, if there is the amplified band of about 650bp size, be then accredited as Foxd1-cre；
DTR-flox double transgenic type mice；
Wherein, in S22, the primer sequence is as follows:
Forward primer F2 5 '-ACCATGAAGCTGCTGCCGTC-3 ', downstream primer R2 5 '-
TCAGTGGGAATTAGTCATGC-3’；
S3, it is accredited as Foxd1-cre from above-mentioned；The embryo of DTR-flox double transgenic type mice separates embryonic kidney, is
Genotype is Foxd1-cre；The double transgenic type embryonic kidney of DTR-flox.
To filter out genotype is Foxd1-cre；The embryonic kidney of DTR-flox double transgenic type shreds that to be inoculated in stem cell special
With in culture medium, in 37 DEG C, 5%CO2Under the conditions of cultivate 72 hours, then in stem cell special culture media add diphtheria toxin, diphtherotoxin
(100ng/ml), liquid (the stem cell special culture media containing 100ng/ml diphtheria toxin, diphtherotoxin) within every 48 hours, is changed until cultivating one week, this
Time remaining cell be Foxd1+Metanephros mesenchymal cell.Unsegregated metanephros mesenchyme is detected thin with Real-time PCR
Born of the same parents and the Foxd1 of extraction+The expression of Foxd1 and Six2 of metanephros mesenchymal cell.Primer sequence is shown in Table 1:
Table 1 Real-time PCR detection primer
Result is as in figure 2 it is shown, Foxd1+Metanephros mesenchymal cell high expressed Foxd1, and express Six2 hardly, thus may be used
Highly purified Foxd1 has been obtained to determine successfully to be extracted by above method+Metanephros mesenchymal cell.
Although, the present invention is described in detail the most with a general description of the specific embodiments, but
On the basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (5)
1. the preparation method of metanephros mesenchymal cell, it is characterised in that by Foxd1creTransgenic mice and DTRfloxTransgenic is little
Mus carries out copulation, separates embryo Ren Mus dirty, culturing embryo nephrocyte, after adding the diphtheria toxin, diphtherotoxin extraction mark Foxd1 positive
Kidney mesenchymal cell.
Method the most according to claim 1, it is characterised in that by Foxd1creTransgenic mice and DTRfloxTransgenic mice
Carry out copulation, obtain female Mus become pregnant start after the embryo in 13.5-15.5 days, separate embryonic kidney, be inoculated in after embryonic kidney is shredded
Cultivating in stem cell special culture media, identify the genotype of each embryo simultaneously, filtering out genotype is Foxd1-
cre；The double transgenic type embryonic kidney of DTR-flox, cultivates addition final concentration 100-150ng/ml in 60-72 hour backward culture medium
Diphtheria toxin, diphtherotoxin, within every 48 hours, change liquid 1 time with the stem cell special culture media containing 100-150ng/ml diphtheria toxin, diphtherotoxin, until cultivate
5-7 days, the survivaling cell collected was the metanephros mesenchymal cell that mark Foxd1 is positive.
Method the most according to claim 2, it is characterised in that described stem cell special culture media has for match industry biotechnology
The C57BL/6 Marrow Mesenchymal Stem Cells complete medium that limit company produces, goods number MUBMX-90011.
The most according to the method in claim 2 or 3, it is characterised in that embryonic kidney is inoculated in stem cell special culture media after shredding
In, in 37 DEG C, 5%CO2Under the conditions of cultivate 2-3 days.
5. according to the method described in any one of claim 2-4, it is characterised in that the genotype of each embryo is identified, sieve
Selecting genotype is Foxd1-cre；Specifically comprising the following steps that of the double transgenic type embryonic kidney of DTR-flox
The qualification of S1, Foxd1-cre transgenic mice
S11, embryo Mus rat-tail extracting genome DNA；
S12, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, 2 ×
EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s, 65 DEG C
30s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S13, result detect: prepare 1.5% agarose gel, and every hole adds PCR product 5 μ l, electrophoresis under 140V voltage
20min, observes electricity kank fruit in gel imaging system, if there is the amplified band of 450bp size, is then accredited as Foxd1-
Cre transgenic mice；
Wherein, in S12, the primer sequence is as follows:
Forward primer F1 5 '-TCTGGTCCAAGAATCCGAAG-3 ', downstream primer R1 5 '-GGGAGGATTGGGAAGACAAT-
3’；
S2、Foxd1-cre；The qualification of DTR-flox double transgenic type mice
S21, it is accredited as the embryo of Foxd1-cre transgenic mice as material with above-mentioned, extracts embryo Mus rat-tail genomic DNA；
S22, employing PCR method detect: PCR reaction system: DNA profiling 2 μ l, 25 μMs of each 0.3 μ l of upstream and downstream primer, 2 ×
EasyTag SuperMix 10 μ l, ddH2O complements to cumulative volume 20 μ l；PCR reaction condition: 94 DEG C of 5min；94 DEG C of 30s, 50 DEG C
45s, 72 DEG C of 30s, repeat 35 circulations；72 DEG C of 10min, 4 DEG C of maintenances；
S23, preparing 1.5% agarose gel, every hole adds PCR product 5 μ l, electrophoresis 20min under 140V voltage, solidifying
Glue imaging system is observed electricity kank fruit, if there is the amplified band of 650bp size, is then accredited as Foxd1-cre；DTR-
Flox double transgenic type mice；
Wherein, in S22, the primer sequence is as follows:
Forward primer F2 5 '-ACCATGAAGCTGCTGCCGTC-3 ', downstream primer R2 5 '-TCAGTGGGAATTAGTCATGC-
3’；
S3, it is accredited as Foxd1-cre from above-mentioned；The embryo of DTR-flox double transgenic type mice separates embryonic kidney, is gene
Type is Foxd1-cre；The double transgenic type embryonic kidney of DTR-flox.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610630153.3A CN106244520B (en) | 2016-08-03 | 2016-08-03 | The preparation method of kidney mesenchymal cell afterwards |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610630153.3A CN106244520B (en) | 2016-08-03 | 2016-08-03 | The preparation method of kidney mesenchymal cell afterwards |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106244520A true CN106244520A (en) | 2016-12-21 |
| CN106244520B CN106244520B (en) | 2019-07-12 |
Family
ID=57607305
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610630153.3A Active CN106244520B (en) | 2016-08-03 | 2016-08-03 | The preparation method of kidney mesenchymal cell afterwards |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106244520B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754728A (en) * | 2016-12-30 | 2017-05-31 | 中国人民解放军总医院 | Metanephros mesenchymal cell system and preparation method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100329986A1 (en) * | 2009-01-06 | 2010-12-30 | Greenbaum Linda E | Adult Hepatic Progenitor Cells and Methods of Use Thereof |
| WO2012037283A2 (en) * | 2010-09-14 | 2012-03-22 | Mount Sinai School Of Medicine | Admimistration of sns neuroprotective agents to promote hematopoietic regeration |
| CN103409448A (en) * | 2013-08-30 | 2013-11-27 | 北京百奥赛图基因生物技术有限公司 | Model mouse preparation method and recombinant vector for conditional cell removal |
-
2016
- 2016-08-03 CN CN201610630153.3A patent/CN106244520B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100329986A1 (en) * | 2009-01-06 | 2010-12-30 | Greenbaum Linda E | Adult Hepatic Progenitor Cells and Methods of Use Thereof |
| WO2012037283A2 (en) * | 2010-09-14 | 2012-03-22 | Mount Sinai School Of Medicine | Admimistration of sns neuroprotective agents to promote hematopoietic regeration |
| CN103409448A (en) * | 2013-08-30 | 2013-11-27 | 北京百奥赛图基因生物技术有限公司 | Model mouse preparation method and recombinant vector for conditional cell removal |
Non-Patent Citations (1)
| Title |
|---|
| JIAN KAN GUO等: "The Terminator mouse is a diphtheria toxin receptor knock in mouse strain for rapid and efficient enrichment of desired cell lineages.", 《KIDNEY INTERNATIONAL》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754728A (en) * | 2016-12-30 | 2017-05-31 | 中国人民解放军总医院 | Metanephros mesenchymal cell system and preparation method and application |
| CN106754728B (en) * | 2016-12-30 | 2020-05-15 | 中国人民解放军总医院 | Metanephric mesenchymal cell line and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106244520B (en) | 2019-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gutierrez-Valdes et al. | Hairy root cultures—a versatile tool with multiple applications | |
| Clevenger et al. | A developmental transcriptome map for allotetraploid Arachis hypogaea | |
| CN107988268A (en) | A kind of method of gene knockout selection and breeding tcf25 Gene Deletion zebra fish | |
| Plett et al. | Endogenous overexpression of Populus MYB186 increases trichome density, improves insect pest resistance, and impacts plant growth | |
| Carman et al. | Apospory appears to accelerate onset of meiosis and sexual embryo sac formation in sorghum ovules | |
| CN105112445A (en) | MiR-205 gene knockout kit based on CRISPR-Cas9 gene knockout technology | |
| CN105492609A (en) | Method for CRISPR-Cas9 specific knockout of pig GGTA1 gene and sgRNA for specific targeted GGTA1 gene | |
| CN105518137A (en) | Method for specifically removing pig SALL1 gene by CRISPR-Cas9 and sgRNA used for specific targeting SALL1 gene | |
| JP2009536830A5 (en) | ||
| CN101849529A (en) | Method for breeding high-quality anti-mite bee variety | |
| Dhekney et al. | Advances in papaya biotechnology | |
| CN103444524B (en) | Method for quickly building genetic transformation regeneration system of grapes | |
| CN106167787A (en) | The preparation of a kind of smoothbark birch xylem protoplast and the method for instantaneous conversion | |
| CN102061297B (en) | Transgenic method for improving salvianolic acid B content in root of red-rooted salvia | |
| Liu et al. | Overexpression of BpCUC2 influences leaf shape and internode development in Betula pendula | |
| JP2021514175A (en) | Method for producing heterogeneous hematopoietic stem cells / progenitor cells using non-mobilized peripheral blood | |
| CN102876713B (en) | Method for improving content of rosmarinic acid and salvianolic acid B in salvia miltiorrhiza bunge while transforming genes | |
| Peng et al. | Backbone phylogeny and adaptive evolution of Pleurospermum sl: New insights from phylogenomic analyses of complete plastome data | |
| CN102465130A (en) | Cloning of XIAO gene for controlling paddy rice strain type, organ size, root and setting percentage property and its application | |
| CN106244520A (en) | The preparation method of metanephros mesenchymal cell | |
| CN104593381B (en) | A kind of corn resistant gene of salt and its application | |
| CN103805576B (en) | Tobacco squalene epoxidase protein, tobacco squalene epoxidase gene and applications of tobacco squalene epoxidase gene | |
| Bellec et al. | New flowering and architecture traits mediated by multiplex CRISPR-Cas9 gene editing in hexaploid Camelina sativa | |
| CN103789325B (en) | Cotton cells wall extensin gene GbEXPATR and application | |
| CN102428963B (en) | Novel applications of corn ZmRop1 protein |
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 |