CN108192860A - A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat - Google Patents

A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat Download PDF

Info

Publication number
CN108192860A
CN108192860A CN201711451315.8A CN201711451315A CN108192860A CN 108192860 A CN108192860 A CN 108192860A CN 201711451315 A CN201711451315 A CN 201711451315A CN 108192860 A CN108192860 A CN 108192860A
Authority
CN
China
Prior art keywords
stem cell
mesenchymal stem
culture medium
fat
amnion mesenchymal
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.)
Pending
Application number
CN201711451315.8A
Other languages
Chinese (zh)
Inventor
熊华强
宋彩霞
孙静
秦连荣
熊惠芳
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.)
Chongqing Sidemu Biological Technology Co Ltd
Original Assignee
Chongqing Sidemu Biological Technology Co Ltd
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 Chongqing Sidemu Biological Technology Co Ltd filed Critical Chongqing Sidemu Biological Technology Co Ltd
Priority to CN201711451315.8A priority Critical patent/CN108192860A/en
Publication of CN108192860A publication Critical patent/CN108192860A/en
Pending legal-status Critical Current

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
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0653—Adipocytes; Adipose tissue
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/32—Amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/33—Insulin
    • 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
    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/13—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells
    • C12N2506/1346—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells from mesenchymal stem cells
    • C12N2506/1392—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells from mesenchymal stem cells from mesenchymal stem cells from other natural sources

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Rheumatology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a kind of culture mediums and abductive approach that amnion mesenchymal stem cell is induced to break up into fat.The culture medium includes basal medium and adding ingredient, wherein, adding ingredient includes the component of following final concentration：0.3 0.6mmol/L of Rosiglitazone, 6 12 μm of ol/L of Pioglitazone, 1 3v/v% of glutamine, 2 6mg/L of 10 20nmol/L of dexamethasone, 100 200nmol/L of insulin and ginsenoside Rg1.Cultural method is using above-mentioned cell culture medium Fiber differentiation amnion mesenchymal stem cell, obtains adipocyte.In culture medium provided by the invention, each component is safe and non-toxic, compatibility is scientific and reasonable, synergistic, and mescenchymal stem cell can be promoted to break up to lipoblast, is substantially shorter divergaence time, improves differentiation efficiency.

Description

A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat
Technical field
The invention belongs to stem cells technology fields, and in particular to a kind of training that amnion mesenchymal stem cell is induced to break up into fat Support base and abductive approach.
Background technology
Mescenchymal stem cell is derived from mesoblastic multipotential stem cell, has height self-renewal capacity and multinomial differentiation Potential.At present in a variety of such as amnions, marrow, umbilical cord, musculature, adipose tissue, bleeding of the umbilicus and dental pulp mature tissue into Work(detaches and turns out mescenchymal stem cell.Amnion is one layer of semi-transparent film in placenta package fetus face, surface without nerve, The tissues such as blood vessel, muscle and lymph can therefrom isolate amniotic epithelial cells and amnion mesenchymal stem cell.Early in more than 50 years Before, scientist and clinicians just have realized that amnion possesses certain biological action.Early stage amnion is once used as covering The dressing of lid skin wound, can the anti-infective, generation that reduces inflammation, and inhibit the immune response of cell, can also stimulate The generation of blood vessel, these features make it be had obtained relatively broad application in the auxiliary treatment of a variety of diseases.
The amnion-derived mescenchymal stem cell of Human plactnta (hAMSCs) comes from the extraembryonic mesoderm of former phase, in certain item There is the potential broken up to multiple cell lines, inside differentiation of germinal layers, such as liver cell, islet-like cells under part；To mesodermal differentiation, Such as osteoblast, cartilage cell, adipocyte, cardiac muscle cell；Outside differentiation of germinal layers, such as nerve cell.Human plactnta amnion comes The mescenchymal stem cell in source not only has the feature of stem cell, also not repellency and immune effect, is cell transplantation and tissue The preferable seed cell of engineering.
Adipocyte, which is applied to beauty, which repairs soft tissue defects, has plasticity strong, is suitable for the excellent of different defect shapes Gesture, the adipogenic induction potential of the amnion-derived mescenchymal stem cell of Human plactnta can become beauty and repair soft tissue defects Source of human stem cell.Therefore, breaking up into fat for the amnion-derived mescenchymal stem cell of Human plactnta is current regenerative medicine and tissue work One of the research hotspot in journey field, adipose tissue regeneration technology are reconstructions and repair fatty as caused by aging and pathological factor etc. Tissue loses and the important technical of damage, and seed cell source of the adipocyte as fat regeneration engineering can be greatly Drive the reconstruction of cambium.Therefore, a kind of method that novel induction amnion mesenchymal stem cell breaks up into fat is researched and developed, is this The technical issues of field technology personnel are urgently to be resolved hurrily.
Invention content
For above-mentioned deficiency of the prior art, the present invention provides a kind of induction amnion mesenchymal stem cells to break up into fat Culture medium and abductive approach, differentiation effect is good, differentiation the period it is short.
To achieve the above object, the technical solution adopted by the present invention to solve the technical problems is：
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, adding ingredient includes the component of following final concentration：Rosiglitazone 0.3-0.6mmol/L, 6-12 μm of ol/L of Pioglitazone, paddy ammonia Amide 1-3v/v%, dexamethasone 10-20nmol/L, insulin 100-200nmol/L and ginsenoside Rg1 2-6mg/L.
Further, adding ingredient includes the component of following final concentration：Rosiglitazone 0.5mmol/L, 10 μ of Pioglitazone Mol/L, glutamine 2v/v%, dexamethasone 15nmol/L, insulin 150nmol/L and ginsenoside Rg1 5mg/L.
Further, basal medium DMEM-H.
Further, 0.5-1% penicillin and 0.5-1% streptomysins are further included in basal medium.
Using above-mentioned culture medium Fiber differentiation amnion mesenchymal stem cell, amnion mesenchymal stem cell used for the third generation extremely 5th generation amnion mesenchymal stem cell, inoculum density are 2-8 × 104A/mL, inducing culturing condition are 37 DEG C, 5%CO2, culture Time is 13-18 days.
The culture medium and abductive approach that induction amnion mesenchymal stem cell provided by the invention breaks up into fat, have with following Beneficial effect：
In culture medium provided by the invention, each component is safe and non-toxic, compatibility is scientific and reasonable, synergistic, is filled between can promoting Matter stem cell breaks up to lipoblast, is substantially shorter divergaence time, improves differentiation efficiency.
Specific embodiment
Embodiment 1
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, basal medium is the DMEM-H containing 0.5% penicillin and 0.5% streptomysin, and adding ingredient includes following final concentration Component：Rosiglitazone 0.3mmol/L, 6 μm of ol/L of Pioglitazone, glutamine 1v/v%, dexamethasone 10nmol/L, insulin 100nmol/L and ginsenoside Rg1 2mg/L.
Embodiment 2
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, basal medium is the DMEM-H containing 0.5% penicillin and 0.5% streptomysin, and adding ingredient includes following final concentration Component：Rosiglitazone 0.6mmol/L, 12 μm of ol/L of Pioglitazone, glutamine 3v/v%, dexamethasone 20nmol/L, pancreas islet Plain 200nmol/L and ginsenoside Rg1 6mg/L.
Embodiment 3
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, basal medium is the DMEM-H containing 0.5% penicillin and 0.5% streptomysin, and adding ingredient includes following final concentration Component：Rosiglitazone 0.4mmol/L, 8 μm of ol/L of Pioglitazone, glutamine 1.5v/v%, dexamethasone 13nmol/L, pancreas islet Plain 130nmol/L and ginsenoside Rg1 3mg/L.
Embodiment 4
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, basal medium is the DMEM-H containing 0.5% penicillin and 0.5% streptomysin, and adding ingredient includes following final concentration Component：Rosiglitazone 0.5mmol/L, 10 μm of ol/L of Pioglitazone, glutamine 2.5v/v%, dexamethasone 18nmol/L, pancreas Island element 180nmol/L and ginsenoside Rg1 4mg/L.
Embodiment 5
It is a kind of to induce the culture medium that breaks up into fat of amnion mesenchymal stem cell, including basal medium and adding ingredient, In, basal medium is the DMEM-H containing 0.5% penicillin and 0.5% streptomysin, and adding ingredient includes following final concentration Component：Rosiglitazone 0.5mmol/L, 10 μm of ol/L of Pioglitazone, glutamine 2v/v%, dexamethasone 15nmol/L, pancreas islet Plain 150nmol/L and ginsenoside Rg1 5mg/L.
Comparative example 1
Compared with Example 5, the difference lies in without ginsenoside for comparative example 1.
Comparative example 2
Compared with Example 5, the difference lies in without dexamethasone for comparative example 2.
Comparative example 3
Compared with Example 5, the difference lies in without dexamethasone and ginsenoside for comparative example 3.
Test example
1st, hAMSCs is separately cultured
(1) 38-40 weeks normal Cesarean esction placenta is aseptically obtained, confirms its source without hepatitis, syphilis, HIV etc. Communicable disease, and endorsed informed consent form with puerpera and its family members.
(2) entire amnion from placenta is sheared off using operating scissors, be then placed in 15cm glass culture dish, remove blood The block position more serious with damage, then rinsed 3-4 times with PBS buffer solution.
(3) amnion is cut into 30-50cm with operating scissors2The fragment of tissue of size, is dispensed into the centrifuge tube of 50mL, wherein The volume of fragment of tissue is 10-15mL.
(4) 0.25% trypsase of fragment of tissue three times volume is added in, is placed in 37 DEG C, in 200r/min constant temperature oscillators 30min is digested, takes out every 10min, acutely rocks.
(5) after digesting, the tissue block digested is transferred to the 250mL liquid storage bottles equipped with PBS buffer solution with tweezers In, PBS solution is added in, after covering tightly lid, acutely shakes, abandons supernatant, so repeat rinsing 2-3 times.
(6) amnion is cut into 10-30cm by the fragment of tissue after step (5) is rinsed using operating scissors2The tissue of size.
(7) the 0.5%I Collagenase Types of 20mL are added in into fragment of tissue obtained by step (6), add DMEM/F12 culture mediums It is 100mL to final volume, is placed in 37 DEG C, digests in 200r/min constant temperature oscillators, acutely rocked with hand every 10min taking-ups, Until tissue block digests completely.
(8) after the completion of digesting, the tissue fluid digested is dispensed into 50mL centrifuge tubes, often pipe 20-25mL, is added in isometric PBS buffer solution, 2000r/min centrifugation 5min.10mLPBS buffer solutions resuspension cell precipitation is added in after abandoning supernatant, with 200 μm of filters Membrane filtration takes filtrate 2000r/min to centrifuge 5min, abandons supernatant, then adds in DMEM/F12 complete mediums of the 5-10mL containing FBS Cell is resuspended, obtains amnion mesenchymal stem cell suspension.
(9) cell count is carried out to cell suspension, cell suspension density is adjusted to 1.0-1.5 × 10 according to count results5 A/mL is inoculated in culture dish, in 5%CO237 DEG C of cultures of incubator.
(10) after cell inoculation 48h, primary cell is carried out to change liquid, it is complete to replace the fresh DMEM/F12 containing 10%FBS Full culture medium places 5%CO237 DEG C of cultures of incubator.
(11) when cell growth to degree of converging is 80-90%, old culture solution is abandoned with suction pipe suction, adds in 2-3mL025% pancreases Protease digests 1-3min, and Microscopic observation cellular contraction becomes bowlder, adds in the DMEM/F12 cultures containing 10%FBS in right amount immediately Liquid terminates digestion, collects cell, and 1500r/min centrifugation 5min abandon supernatant, add in complete containing the DMEM/F12 containing 10%FBS in right amount Full culture medium carries out cell count, by 1.0 × 105A/mL density, which is inoculated in culture dish, carries out secondary culture, is placed in 5%CO2 37 DEG C of cultures of incubator, it is primary every passage in 3-4 days.
2nd, to being in the 2nd generation amnion mesenchymal stem cell of exponential phase, using Flow cytometry surface, it is anti- Former expression, then analysis result, the result shows that obtained hAMSCs derives from mesoblastic mescenchymal stem cell.
3rd, amnion mesenchymal stem cell into fat break up
Take P3 for amnion mesenchymal stem cell by 2 × 10 respectively4A/mL density is inoculated in basal medium and cultivates, training Support to cell fusion degree reach more than 90% when, remove old culture medium, add in embodiment 1-5 and comparative example 1-3 culture mediums, It is subsequently placed in 5%CO237 DEG C of cultures of incubator, change the liquid once for every 3 days.
After cell culture to after 13-18 days, cell culture supernatant is drawn, then clean cell 2-3 times with PBS, then plus Enter 4% paraformaldehyde and carry out cell fixation 15-20min, suction abandons paraformaldehyde, oil red O stain liquid is added in after cleaning 3 times with PBS It is dyed, induction differentiation rate is calculated according to staining conditions.
Induce differentiation rate=staining positive cells number/total number of cells × 100%
As a result：After culture 13-18 days, the induction differentiation efficiency of embodiment 1-5 is significantly higher than comparative example 1-3, especially Embodiment 5 induces differentiation rate highest, illustrates the culture medium for preparing of the present invention to amnion mesenchymal stem cell into fat induction Effect is very good, it can in a short time broken up into fat, differentiation rate is induced to be up to 65.9% He respectively at 13 days and 18 days 95.7%.

Claims (8)

1. a kind of culture medium that amnion mesenchymal stem cell is induced to break up into fat, which is characterized in that including basal medium and add Addition point, wherein, adding ingredient includes the component of following final concentration：Rosiglitazone 0.3-0.6mmol/L, Pioglitazone 6-12 μ Mol/L, glutamine 1-3v/v%, dexamethasone 10-20nmol/L, insulin 100-200nmol/L and ginsenoside Rg1 2- 6mg/L。
2. the culture medium that induction amnion mesenchymal stem cell according to claim 1 breaks up into fat, which is characterized in that described Adding ingredient includes the component of following final concentration：Rosiglitazone 0.5mmol/L, 10 μm of ol/L of Pioglitazone, glutamine 2v/ V%, dexamethasone 15nmol/L, insulin 150nmol/L and ginsenoside Rg1 5mg/L.
3. the culture medium that induction amnion mesenchymal stem cell according to claim 1 or 2 breaks up into fat, which is characterized in that The basal medium is DMEM-H.
4. the culture medium that induction amnion mesenchymal stem cell according to claim 3 breaks up into fat, which is characterized in that described 0.5-1% penicillin and 0.5-1% streptomysins are further included in basal medium.
A kind of 5. method that amnion mesenchymal stem cell is induced to break up into fat, which is characterized in that using any one of claim 1-4 The cell culture medium Fiber differentiation amnion mesenchymal stem cell obtains adipocyte.
6. according to the method described in claim 5, it is characterized in that, the amnion mesenchymal stem cell is the third generation to the 5th generation Amnion mesenchymal stem cell.
7. according to the method described in claim 5, it is characterized in that, the inducing culturing condition be 37 DEG C, 5%CO2。
8. according to the method described in claim 5, it is characterized in that, the Fiber differentiation time is 13-18 days.
CN201711451315.8A 2017-12-27 2017-12-27 A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat Pending CN108192860A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711451315.8A CN108192860A (en) 2017-12-27 2017-12-27 A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711451315.8A CN108192860A (en) 2017-12-27 2017-12-27 A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat

Publications (1)

Publication Number Publication Date
CN108192860A true CN108192860A (en) 2018-06-22

Family

ID=62584774

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711451315.8A Pending CN108192860A (en) 2017-12-27 2017-12-27 A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat

Country Status (1)

Country Link
CN (1) CN108192860A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949676A (en) * 2018-07-03 2018-12-07 湖南未名三胞转化医学科技有限公司 A kind of ginsenoside Rg1's autologous fat stem cell extracorporeal culturing method
CN111979185A (en) * 2020-09-08 2020-11-24 依科赛生物科技（太仓）有限公司 Mesenchymal stem cell adipogenic differentiation medium and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107460163A (en) * 2017-05-08 2017-12-12 广州赛莱拉干细胞科技股份有限公司 A kind of method that cell culture medium breaks up with induction amnion mesenchymal stem cell into fat

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107460163A (en) * 2017-05-08 2017-12-12 广州赛莱拉干细胞科技股份有限公司 A kind of method that cell culture medium breaks up with induction amnion mesenchymal stem cell into fat

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
EUN-JEONG KOH 等: "Ginsenoside Rg1 suppresses early stage of adipocyte development via activation of C/EBP homologous protein-10 in 3T3-L1 and attenuates fat accumulation in high fat diet-induced obese zebrafish", 《J GINSENG RES》 *
李烨: "间充质干细胞羰基应激及人参皂苷抗羰基应激作用", 《万方》 *
王爽: "人参皂苷Rg1对体外培养人脐带血间充质干细胞衰老的影响", 《万方》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949676A (en) * 2018-07-03 2018-12-07 湖南未名三胞转化医学科技有限公司 A kind of ginsenoside Rg1's autologous fat stem cell extracorporeal culturing method
CN111979185A (en) * 2020-09-08 2020-11-24 依科赛生物科技（太仓）有限公司 Mesenchymal stem cell adipogenic differentiation medium and application thereof

Similar Documents

Publication Publication Date Title
CN104263697B (en) A kind of method that inducing culture and induction human adipose mesenchymal stem cells generate insulin secretory cell
CN103805562B (en) Cultivate the serum free medium of placenta mesenchyma stem cell
CN101914490B (en) Human amnion mesenchymal stem cell serum-free culture medium and culture method thereof
CN102127522B (en) Human umbilical mesenchymal stem cell and preparation method thereof
CN102676452B (en) Culture medium containing human umbilical cord mesenchymal stem cell exudates and preparation method and applications thereof
CN104450611A (en) Primary separation and culture method of human amniotic mesenchymal stem cells
CN104726406B (en) It is a kind of to induce the method that dental pulp Derived from Mesenchymal Stem Cells is nerve cell
CN105238750B (en) A kind of method of recovery umbilical cord mesenchymal stem cells
CN105713871A (en) Human chorion mesenchymal stem cell isolated culture method
CN109234229B (en) Method for separating mesenchymal stem cells from placental blood vessels and digestive enzyme composition used in same
CN103191154B (en) Mesenchymal stem cells and the application of extracting method in the psoriatic medicine of preparation thereof
CN106085952A (en) A kind of placental villi plate mescenchymal stem cell and extracting method thereof
CN103966159B (en) Human plactnta Subaerial blue green algae and stem cell bank construction process thereof
CN104818244A (en) Amnion epithelial cell separation and culture method
CN105670987A (en) Inhibition method for induced differentiation of hair follicle stem cells into vascular endothelial cells
CN108192860A (en) A kind of culture medium and abductive approach that amnion mesenchymal stem cell is induced to break up into fat
CN107460163A (en) A kind of method that cell culture medium breaks up with induction amnion mesenchymal stem cell into fat
CN104762257A (en) Method for preparing mesenchymal stem cell from umbilical cord
CN106754685A (en) A kind of construction method of Human fat mesenchymal stem cell bank
CN106244533A (en) The primary separation method of gingiva mescenchymal stem cell
CN107937346A (en) A kind of method by the use of human urine cell as feeder layer culture induced multi-potent stem cell
CN108373990B (en) Culture medium for inducing differentiation from human amniotic epithelial stem cells to melanocytes and method thereof
CN109628388B (en) Isolation of mesenchymal stem cells from placental blood vessels with digestive enzyme composition
CN105112359A (en) Separating and cultivating method of mouse umbilical cord mesenchymal stem cells
CN104031881B (en) A kind of isolated culture method of mankind's olfactory mucosa mescenchymal stem cell

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication

Application publication date: 20180622

RJ01 Rejection of invention patent application after publication