CN106884004A - A kind of preparation method of efficient fell skin tissue multipotential stem cell - Google Patents
A kind of preparation method of efficient fell skin tissue multipotential stem cell Download PDFInfo
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- CN106884004A CN106884004A CN201710176129.1A CN201710176129A CN106884004A CN 106884004 A CN106884004 A CN 106884004A CN 201710176129 A CN201710176129 A CN 201710176129A CN 106884004 A CN106884004 A CN 106884004A
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- 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
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
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- 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
Abstract
The invention discloses a kind of preparation method of efficient fell skin tissue multipotential stem cell, concretely comprise the following steps：It is prepared by coating bottle：Coating bottle is saved backup in putting 4 DEG C of refrigerators；It is prepared by skin progenitor cell：Cheek or chin skin histology are taken, adipose tissue is removed；Tissue block is placed in sterilized petri dishes, digested overnight；Skin corium fritter is digested；Above-mentioned cell suspension is filtered with filter；Continue to employ attached cell；Cultivated in incubator；Bottom of bottle is blown and beaten repeatedly with suction pipe, makes cell detachment；Above-mentioned cell is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reached for 25 generations, you can.The present invention has the advantages that cell yield is high, differentiation efficiency is high.
Description
Technical field
The present invention relates to cell engineering field, specifically a kind of preparation of efficient fell skin tissue multipotential stem cell
Method.
Background technology
The multipotential stem cell for coming from adult tissue is the important sources of organizational project and regenerative medicine field, and it is easy to obtain
, and with extensive medical applications prospect.Although embryonic pleuripotent stem cell has great potentiality in regenerative medicine field,
But due to the ethics problem of heterogenous cell transplanting, make its should be restricted clinically.Induced multi-potent stem cell (iPS) is
Another is potentially originated, but iPS cells have potential carcinogenicity, it is become difficulty in clinical practice.
Epidermal stem cells are skin histology specific stem cells, with potential versatility.Its ectoderm for deriving from embryo,
There is more original differentiation capability, the feature with non-mature cell, cell volume is small, and organelle is few, remains static, mainly
Positioned at the basal cell layer of skin, and raw coal bunker knuckle portion.It has versatility, can be divided into nerve cell, fat
The histocytes such as cell, cartilage cell.Epidermal stem cells have important grinding in gene therapy, cell therapy, organizational project
Study carefully and application value.Research research on epidermal stem cells has become life science and organizational project, regenerative medicine field
One of forward position of research.
Although skin progenitor cell is a kind of important source, however, with advancing age, the stem cell in application on human skin
Reduce therewith, its differentiation capability also gradually weakens.Recently, studies have found that, before the skin source property of people's cheek or chin
Compared with people's eyelid, four limbs, the skin precursor stem cell of trunk, it has more significant Stem Cell Activity to body cell, with more
Differentiation efficiency high, this also show people's cheek and chin skin as one important stem cell of organizational project and regenerative medicine
Source, with energetically application prospect.
The content of the invention
The invention aims to solve the defect that cell yield of the prior art is low, differentiation efficiency is low, there is provided one
The preparation method of efficient fell skin tissue multipotential stem cell is planted to solve the above problems.
The invention discloses a kind of preparation method of efficient fell skin tissue multipotential stem cell, concretely comprise the following steps：
First, prepared by coating bottle：
(1), people's type Ⅳ collagen is dissolved and is diluted to 0.1g/L with PBS and is made coating buffer；
(2), take in 5ml coating buffers addition blake bottle, keep flat, put 4 DEG C of refrigerator overnights；
(3) supernatant, is abandoned, is placed in superclean bench and is dried；
(4), coating bottle is saved backup in putting 4 DEG C of refrigerators；
2nd, prepared by skin progenitor cell：
(5) cheek or chin skin histology, are taken, adipose tissue is removed；
(6), skin be cut into the fritter of 0.5mm × 0.5mm, flushed three times with PBS；
(7), tissue block is placed in sterilized petri dishes, and corium faces down, with the digestive juice containing 1U/ml neutral proteinases II, 4
DEG C, digested overnight；
(8) epidermis, is separated with ophthalmology tweezers, and abandons it, skin corium is then cut into 1mm3Fritter；
(9), with containing the digestive juice that volume fraction is 0.5% NTx enzyme, in 37 DEG C, skin corium fritter jog is digested
1h；
(10) the DMEM/F12 culture mediums of equivalent, are added, terminates digestion, then cell suspension is centrifuged 10min with 180g；
(11) above-mentioned cell suspension is filtered with filter, removes remaining tissue block, adjustment cell density is 1 × 105/
ml；
(12), by above-mentioned cell suspension inoculation in the T25 blake bottles for being coated with type Ⅳ collagen, 37 DEG C, 5%CO are placed in2
In incubator, after adherent 10-20min, supernatant is abandoned in suction, continues to employ attached cell；
(13) 37 DEG C, 5%CO, are placed in 10ml cell culture fluids are added in T25 blake bottles again2Saturated humidity training
Culture in case is supported, it is every to change within 2-3 days liquid once；
(14) after, cell fusion degree reaches more than 80%, nutrient solution is blotted, cell is rinsed twice with PBS liquid；
(15) it is 0.25% tryptic digestive juice+0.01%EDTA that volume fraction, is added in blake bottle, is placed in 37 DEG C,
5% CO2Cell retraction, space between cells increase are observed after incubator 2-3min, under inverted microscope, then is added in blake bottle
10ml cell culture fluids terminate digestion；
(16) bottom of bottle, is blown and beaten repeatedly with suction pipe, makes cell detachment, then cell suspension 180g is centrifuged, 5min；
(17) above-mentioned cell, is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reaches 2-5 generations, you can.
Preferably, in described step (7), described digestive juice is DMEM/F12 culture mediums, described DMEM/F12
Contain 1U/ml neutral proteinases II, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs in culture medium.
Preferably, in described step (9), described digestive juice uses DMEM/F12 culture mediums, described DMEM/
It is 0.5% NTx enzyme, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs to contain volume fraction in F12 culture mediums.
Preferably, in described step (11), the aperture of described filter is 40 μm.
Preferably, in described step (13), described cell culture fluid is DMEM/F12 culture mediums, described
Contain B-27,20ng/ml EGF, 40ng/ml bFGF, 40U/ml sulfuric acid celebrating that volume fraction is 2% in DMEM/F12 culture mediums
Big mycin, 25 μ g/ml amphotericin Bs.
The present invention has advantages below compared to existing technology：
1st, the present invention selects the postoperative people's cheek of surgical plastic or chin skin as the source of stem cell, and its stem cell lives
Property it is strong, differentiation efficiency is high；
2nd, using the blake bottle inoculating cell for being coated with type Ⅳ collagen, it is conducive to keeping stem cell proterties the present invention, also has
Beneficial to promotion stem cells hyperplasia；
3rd, using the blake bottle inoculating cell for being coated with type Ⅳ collagen, it is 10- to control the cell attachment time to the present invention
20min, can screen the skin derived stem cells for obtaining high-purity.
Specific embodiment
The invention discloses a kind of preparation method of efficient fell skin tissue multipotential stem cell, concretely comprise the following steps：
First, prepared by coating bottle：
(1), people's type Ⅳ collagen is dissolved and is diluted to 0.1g/L with PBS and is made coating buffer；
(2), take in 5ml coating buffers addition blake bottle, keep flat, put 4 DEG C of refrigerator overnights；
(3) supernatant, is abandoned, is placed in superclean bench and is dried；
(4), coating bottle is saved backup in putting 4 DEG C of refrigerators；
2nd, prepared by skin progenitor cell：
(5) cheek or chin skin histology, are taken, adipose tissue is removed；
(6), skin be cut into the fritter of 0.5mm × 0.5mm, flushed three times with PBS；
(7), tissue block is placed in sterilized petri dishes, and corium faces down, with the digestive juice containing 1U/ml neutral proteinases II, 4
DEG C, digested overnight；
(8) epidermis, is separated with ophthalmology tweezers, and abandons it, skin corium is then cut into 1mm3Fritter；
(9), with containing the digestive juice that volume fraction is 0.5% NTx enzyme, in 35 DEG C, skin corium fritter jog is digested
1h；
(10) the DMEM/F12 culture mediums of equivalent, are added, terminates digestion, then cell suspension is centrifuged 10min with 180g；
(11) above-mentioned cell suspension is filtered with filter, removes remaining tissue block, adjustment cell density is 1 × 105/
ml；
(12), by above-mentioned cell suspension inoculation in the T25 blake bottles for being coated with type Ⅳ collagen, 37 DEG C, 5%CO are placed in2
In incubator, after adherent 10-20min, supernatant is abandoned in suction, continues to employ attached cell；
(13) 37 DEG C, 5%CO, are placed in 10ml cell culture fluids are added in T25 blake bottles again2Saturated humidity training
Culture in case is supported, it is every to change within 2-3 days liquid once；
(14) after, cell fusion degree reaches more than 80%, nutrient solution is blotted, cell is rinsed twice with PBS liquid；
(15) it is 0.25% tryptic digestive juice+0.01%EDTA that volume fraction, is added in blake bottle, is placed in 37 DEG C,
5% CO2Cell retraction, space between cells increase are observed after incubator 2-3min, under inverted microscope, then is added in blake bottle
10ml cell culture fluids terminate digestion；
(16) bottom of bottle, is blown and beaten repeatedly with suction pipe, makes cell detachment, then cell suspension 180g is centrifuged, 5min；
(17) above-mentioned cell, is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reaches 2-5 generations, you can.
Preferably, in described step (7), described digestive juice is DMEM/F12 culture mediums, described DMEM/F12
Contain 1U/ml neutral proteinases II, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs in culture medium.
Preferably, in described step (9), described digestive juice uses DMEM/F12 culture mediums, described DMEM/
It is 0.5% NTx enzyme, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs to contain volume fraction in F12 culture mediums.
Preferably, in described step (11), the aperture of described filter is 40 μm.
Preferably, in described step (13), described cell culture fluid is DMEM/F12 culture mediums, described
Contain B-27,20ng/ml EGF, 40ng/ml bFGF, 40U/ml sulfuric acid celebrating that volume fraction is 2% in DMEM/F12 culture mediums
Big mycin, 25 μ g/ml amphotericin Bs.
Embodiment 1
The invention discloses a kind of preparation method of efficient fell skin tissue multipotential stem cell, concretely comprise the following steps：
First, prepared by coating bottle：
(1), people's type Ⅳ collagen is dissolved and is diluted to 0.1g/L with PBS and is made coating buffer；
(2), take in 5ml coating buffers addition blake bottle, keep flat, put 4 DEG C of refrigerator overnights；
(3) supernatant, is abandoned, is placed in superclean bench and is dried；
(4), coating bottle is saved backup in putting 4 DEG C of refrigerators；
2nd, prepared by skin progenitor cell：
(5) cheek tissue, is taken, adipose tissue is removed；
(6), skin be cut into the fritter of 0.5mm × 0.5mm, flushed three times with PBS；
(7), tissue block is placed in sterilized petri dishes, and corium faces down, with the digestive juice containing 1U/ml neutral proteinases II, 4
DEG C, digested overnight；
(8) epidermis, is separated with ophthalmology tweezers, and abandons it, skin corium is then cut into 1mm3Fritter；
(9), with containing the digestive juice that volume fraction is 0.5% NTx enzyme, in 35 DEG C, skin corium fritter jog is digested
1h；
(10) the DMEM/F12 culture mediums of equivalent, are added, terminates digestion, then cell suspension is centrifuged 10min with 180g；
(11) above-mentioned cell suspension is filtered with filter, removes remaining tissue block, adjustment cell density is 1 × 105/
ml；
(12), by above-mentioned cell suspension inoculation in the T25 blake bottles for being coated with type Ⅳ collagen, 37 DEG C, 5%CO are placed in2
In incubator, after adherent 15min, supernatant is abandoned in suction, continues to employ attached cell；
(13) 37 DEG C, 5%CO, are placed in 10ml cell culture fluids are added in T25 blake bottles again2Saturated humidity training
Culture in case is supported, liquid is changed once within every 2 days；
(14) after, cell fusion degree reaches more than 80%, nutrient solution is blotted, cell is rinsed twice with PBS liquid；
(15) it is 0.25% tryptic digestive juice+0.01%EDTA that volume fraction, is added in blake bottle, is placed in 37 DEG C,
5% CO2Cell retraction, space between cells increase are observed after incubator 2min, under inverted microscope, then is added in blake bottle
10ml cell culture fluids terminate digestion；
(16) bottom of bottle, is blown and beaten repeatedly with suction pipe, makes cell detachment, then cell suspension 180g is centrifuged, 5min；
(17) above-mentioned cell, is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reached for 3 generations, you can.
In described step (7), described digestive juice is DMEM/F12 culture mediums, is contained in described DMEM/F12 culture mediums
There are 1U/ml neutral proteinases II, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs.
In described step (9), described digestive juice uses DMEM/F12 culture mediums, in described DMEM/F12 culture mediums
It is 0.5% NTx enzyme, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs containing volume fraction.
In described step (11), the aperture of described filter is 40 μm.
In described step (13), described cell culture fluid is DMEM/F12 culture mediums, described DMEM/F12 cultures
Contain B-27,20ng/ml EGF, 40ng/ml bFGF, 40U/ml gentamicin sulphate, the 25 μ g/ that volume fraction is 2% in base
Ml amphotericin Bs.
Embodiment 2
The invention discloses a kind of preparation method of efficient fell skin tissue multipotential stem cell, concretely comprise the following steps：
First, prepared by coating bottle：
(1), people's type Ⅳ collagen is dissolved and is diluted to 0.1g/L with PBS and is made coating buffer；
(2), take in 5ml coating buffers addition blake bottle, keep flat, put 4 DEG C of refrigerator overnights；
(3) supernatant, is abandoned, is placed in superclean bench and is dried；
(4), coating bottle is saved backup in putting 4 DEG C of refrigerators；
2nd, prepared by skin progenitor cell：
(5) bar skin histology, is removed, adipose tissue is removed；
(6), skin be cut into the fritter of 0.5mm × 0.5mm, flushed three times with PBS；
(7), tissue block is placed in sterilized petri dishes, and corium faces down, with the digestive juice containing 1U/ml neutral proteinases II, 4
DEG C, digested overnight；
(8) epidermis, is separated with ophthalmology tweezers, and abandons it, skin corium is then cut into 1mm3Fritter；
(9), with containing the digestive juice that volume fraction is 0.5% NTx enzyme, in 35 DEG C, skin corium fritter jog is digested
1h；
(10) the DMEM/F12 culture mediums of equivalent, are added, terminates digestion, then cell suspension is centrifuged 10min with 180g；
(11) above-mentioned cell suspension is filtered with filter, removes remaining tissue block, adjustment cell density is 1 × 105/
ml；
(12), by above-mentioned cell suspension inoculation in the T25 blake bottles for being coated with type Ⅳ collagen, 37 DEG C, 5%CO are placed in2
In incubator, after adherent 20min, supernatant is abandoned in suction, continues to employ attached cell；
(13) 37 DEG C, 5%CO, are placed in 10ml cell culture fluids are added in T25 blake bottles again2Saturated humidity training
Culture in case is supported, liquid is changed once within every 2 days；
(14) after, cell fusion degree reaches more than 80%, nutrient solution is blotted, cell is rinsed twice with PBS liquid；
(15) it is 0.25% tryptic digestive juice+0.01%EDTA that volume fraction, is added in blake bottle, is placed in 37 DEG C,
5% CO2Cell retraction, space between cells increase are observed after incubator 3min, under inverted microscope, then is added in blake bottle
10ml cell culture fluids terminate digestion；
(16) bottom of bottle, is blown and beaten repeatedly with suction pipe, makes cell detachment, then cell suspension 180g is centrifuged, 5min；
(17) above-mentioned cell, is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reached for 2 generations, you can.
In described step (7), described digestive juice is DMEM/F12 culture mediums, in described DMEM/F12 culture mediums
Contain 1U/ml neutral proteinases II, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs.
In described step (9), described digestive juice uses DMEM/F12 culture mediums, in described DMEM/F12 culture mediums
It is 0.5% NTx enzyme, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs containing volume fraction.
In described step (11), the aperture of described filter is 40 μm.
In described step (13), described cell culture fluid is DMEM/F12 culture mediums, described DMEM/F12 cultures
Contain B-27,20ng/ml EGF, 40ng/ml bFGF, 40U/ml gentamicin sulphate, the 25 μ g/ that volume fraction is 2% in base
Ml amphotericin Bs.
General principle of the invention, principal character and advantages of the present invention has been shown and described above.The technology of the industry
Personnel it should be appreciated that the present invention is not limited to the above embodiments, the simply present invention described in above-described embodiment and specification
Principle, various changes and modifications of the present invention are possible without departing from the spirit and scope of the present invention, these change and
Improvement is both fallen within the range of claimed invention.The protection domain of application claims by appending claims and its
Equivalent is defined.
Claims (5)
1. a kind of preparation method of efficient fell skin tissue multipotential stem cell, it is characterised in that：Concretely comprise the following steps：
First, prepared by coating bottle：
(1), people's type Ⅳ collagen is dissolved and is diluted to 0.1g/L with PBS and is made coating buffer；
(2), take in 5ml coating buffers addition blake bottle, keep flat, put 4 DEG C of refrigerator overnights；
(3) supernatant, is abandoned, is placed in superclean bench and is dried；
(4), coating bottle is saved backup in putting 4 DEG C of refrigerators；
2nd, prepared by skin progenitor cell：
(5) cheek or chin skin histology, are taken, adipose tissue is removed；
(6), skin be cut into the fritter of 0.5mm × 0.5mm, flushed three times with PBS；
(7), tissue block is placed in sterilized petri dishes, and corium faces down, with the digestive juice containing 1U/ml neutral proteinases II, 4 DEG C,
Digested overnight；
(8) epidermis, is separated with ophthalmology tweezers, and abandons it, skin corium is then cut into 1mm3Fritter；
(9), with containing the digestive juice that volume fraction is 0.5% NTx enzyme, in 37 DEG C, 1h is digested to skin corium fritter jog；
(10) the DMEM/F12 culture mediums of equivalent, are added, terminates digestion, then cell suspension is centrifuged 10min with 180g；
(11) above-mentioned cell suspension is filtered with filter, removes remaining tissue block, adjustment cell density is 1 × 105/ml；
(12), by above-mentioned cell suspension inoculation in the T25 blake bottles for being coated with type Ⅳ collagen, 37 DEG C, 5%CO are placed in2Incubator
In, after adherent 10-20min, supernatant is abandoned in suction, continues to employ attached cell；
(13) 37 DEG C, 5%CO, are placed in 10ml cell culture fluids are added in T25 blake bottles again2Saturated humidity incubator in
Culture, it is every to change within 2-3 days liquid once；
(14) after, cell fusion degree reaches more than 80%, nutrient solution is blotted, cell is rinsed twice with PBS liquid；
(15) it is 0.25% tryptic digestive juice+0.01%EDTA that volume fraction, is added in blake bottle, is placed in 37 DEG C, 5%
CO2Observe cell retraction after incubator 2-3min, under inverted microscope, space between cells increase, then to adding 10ml in blake bottle
Cell culture fluid terminates digestion；
(16) bottom of bottle, is blown and beaten repeatedly with suction pipe, makes cell detachment, then cell suspension 180g is centrifuged, 5min；
(17) above-mentioned cell, is pressed 1:3 ratio sub-bottle passage, continues to cultivate, and cell reaches 2-5 generations, you can.
2. the preparation method of a kind of efficient fell skin tissue multipotential stem cell according to claim 1, it is characterised in that：
In described step (7), described digestive juice is DMEM/F12 culture mediums, and 1U/ml is contained in described DMEM/F12 culture mediums
Neutral proteinase II, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs.
3. the preparation method of a kind of efficient fell skin tissue multipotential stem cell according to claim 1, it is characterised in that：
In described step (9), described digestive juice uses DMEM/F12 culture mediums, and volume is contained in described DMEM/F12 culture mediums
Fraction is 0.5% NTx enzyme, 40U/ml gentamicin sulphates, 25 μ g/ml amphotericin Bs.
4. the preparation method of a kind of efficient fell skin tissue multipotential stem cell according to claim 1, it is characterised in that：
In described step (11), the aperture of described filter is 40 μm.
5. the preparation method of a kind of efficient fell skin tissue multipotential stem cell according to claim 1, it is characterised in that：
In described step (13), described cell culture fluid is DMEM/F12 culture mediums, is contained in described DMEM/F12 culture mediums
Volume fraction be 2% B-27,20ng/ml EGF, 40ng/mlbFGF, 40U/ml gentamicin sulphate, 25 μ g/ml both sexes it is mould
Plain B.
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| CN201710176129.1A CN106884004A (en) | 2017-03-23 | 2017-03-23 | A kind of preparation method of efficient fell skin tissue multipotential stem cell |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110387348A (en) * | 2018-04-18 | 2019-10-29 | 江苏齐氏生物科技有限公司 | A kind of isolation and culture method of application on human skin cutin cambial cell |
| CN113957038A (en) * | 2021-10-22 | 2022-01-21 | 青岛农业大学 | In-vitro separation method of human skin stem cells |
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|---|---|---|---|---|
| CN103865872A (en) * | 2013-12-10 | 2014-06-18 | 中国人民解放军第三军医大学第一附属医院 | Method for separating and culturing human epidermal stem cells |
| CN104450856A (en) * | 2014-12-04 | 2015-03-25 | 中国检验检疫科学研究院 | Application of epidermal stem cells in detecting cytotoxicity of cosmetics and raw materials of cosmetics |
| CN105062959A (en) * | 2015-09-20 | 2015-11-18 | 领航干细胞再生医学工程有限公司 | Isolated culture method of human amnia mesenchymal stem cells |
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2017
- 2017-03-23 CN CN201710176129.1A patent/CN106884004A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103865872A (en) * | 2013-12-10 | 2014-06-18 | 中国人民解放军第三军医大学第一附属医院 | Method for separating and culturing human epidermal stem cells |
| CN104450856A (en) * | 2014-12-04 | 2015-03-25 | 中国检验检疫科学研究院 | Application of epidermal stem cells in detecting cytotoxicity of cosmetics and raw materials of cosmetics |
| CN105062959A (en) * | 2015-09-20 | 2015-11-18 | 领航干细胞再生医学工程有限公司 | Isolated culture method of human amnia mesenchymal stem cells |
Non-Patent Citations (3)
| Title |
|---|
| KATSUHIRO YOSHIKAWA等: ""Multipotent stem cells are effectively collected from adult human cheek skin"", 《BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS》 * |
| 杨丽等: ""人皮肤表皮干细胞体外培养与应用研究进展"", 《现代生物医学进展》 * |
| 林才等: ""人表皮干细胞的快速分离培养基鉴定"", 《上海交通大学学报（医学版）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110387348A (en) * | 2018-04-18 | 2019-10-29 | 江苏齐氏生物科技有限公司 | A kind of isolation and culture method of application on human skin cutin cambial cell |
| CN113957038A (en) * | 2021-10-22 | 2022-01-21 | 青岛农业大学 | In-vitro separation method of human skin stem cells |
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