CN113755433A - Synovial mesenchymal stem cell suspension and preparation method thereof - Google Patents
Synovial mesenchymal stem cell suspension and preparation method thereof Download PDFInfo
- Publication number
- CN113755433A CN113755433A CN202110907400.0A CN202110907400A CN113755433A CN 113755433 A CN113755433 A CN 113755433A CN 202110907400 A CN202110907400 A CN 202110907400A CN 113755433 A CN113755433 A CN 113755433A
- Authority
- CN
- China
- Prior art keywords
- mesenchymal stem
- synovial
- culture
- suspension
- stem cells
- 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
Links
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 151
- 239000006285 cell suspension Substances 0.000 title claims abstract description 75
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 210000004027 cell Anatomy 0.000 claims abstract description 106
- 238000000034 method Methods 0.000 claims abstract description 62
- 210000001258 synovial membrane Anatomy 0.000 claims abstract description 48
- 239000000725 suspension Substances 0.000 claims abstract description 27
- 230000003321 amplification Effects 0.000 claims abstract description 18
- 238000012258 culturing Methods 0.000 claims abstract description 18
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 18
- 238000004806 packaging method and process Methods 0.000 claims abstract description 9
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 57
- 239000001963 growth medium Substances 0.000 claims description 49
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 48
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 32
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 24
- 239000001569 carbon dioxide Substances 0.000 claims description 24
- 239000012879 subculture medium Substances 0.000 claims description 24
- 239000002609 medium Substances 0.000 claims description 23
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 16
- 229930182555 Penicillin Natural products 0.000 claims description 16
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 16
- 239000012091 fetal bovine serum Substances 0.000 claims description 16
- 229940049954 penicillin Drugs 0.000 claims description 16
- 229960005322 streptomycin Drugs 0.000 claims description 16
- 102000004142 Trypsin Human genes 0.000 claims description 15
- 108090000631 Trypsin Proteins 0.000 claims description 15
- 239000012588 trypsin Substances 0.000 claims description 15
- 239000000835 fiber Substances 0.000 claims description 12
- 239000003102 growth factor Substances 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 11
- 230000029087 digestion Effects 0.000 claims description 9
- 238000005516 engineering process Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 9
- 239000002504 physiological saline solution Substances 0.000 claims description 9
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 8
- 238000012136 culture method Methods 0.000 claims description 8
- 229920002674 hyaluronan Polymers 0.000 claims description 8
- 229960003160 hyaluronic acid Drugs 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 8
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 claims description 7
- 230000004927 fusion Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000004246 zinc acetate Substances 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 230000000694 effects Effects 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 206010007710 Cartilage injury Diseases 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 101710151387 Serine protease 1 Proteins 0.000 description 1
- 102100032491 Serine protease 1 Human genes 0.000 description 1
- 101710119665 Trypsin-1 Proteins 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 230000009816 chondrogenic differentiation Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000107 myocyte Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
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
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/119—Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
-
- 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
- C12N2501/90—Polysaccharides
- C12N2501/905—Hyaluronic acid
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (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 synovial membrane mesenchymal stem cell suspension and a preparation method thereof, belonging to the technical field of medical manufacturing. It comprises the following steps: (1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell; (2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture; (3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell. The preparation method of the synovial mesenchymal stem cell suspension is simple in process and easy to manufacture, and is beneficial to improving the suspension stability of the synovial mesenchymal stem cells.
Description
Technical Field
The invention belongs to the technical field of medical manufacturing, and particularly relates to a synovial mesenchymal stem cell suspension and a preparation method thereof.
Background
Mesenchymal Stem Cells (MSCs) refer to clonal cells derived from mesenchymal tissue with high proliferative and multipotent differentiation potential. A large number of researches prove that the MSCs have the directional differentiation capacity to adipocytes, osteoblasts, chondrocytes, myocytes and the like, and increasingly show wide application prospects in the fields of tissue engineering, regenerative medicine and the like.
And is considered to be the most ideal seed cell for repairing cartilage damage due to the excellent chondrogenic differentiation potential. However, the related studies of mesenchymal stem cells are relatively rare and the biological characteristics and possible effects are poorly understood. Currently, synovial mesenchymal stem cells of mice can be used for simulation experiment research. For example, the Chinese patent, application number: cn201810155580.x, publication No.: CN108456655A discloses a mesenchymal stem cell suspension, a preparation method and an application thereof, and the technical proposal is as follows: obtaining a primary mesenchymal stem cell; culturing and amplifying the primary mesenchymal stem cell: after 5-10 days of culture of the primary mesenchymal stem cells, the primary mesenchymal stem cells are replaced by serum-free DMEM nutrient solution until 65-75% of the cells are fused, culture supernatant is removed, first digestive fluid is added, the cells are digested for 0.5-2min, the cells are added into the removed culture supernatant after being contracted and then are neutralized for centrifugal separation, complete culture medium is added again for heavy suspension, inoculation is carried out according to the 1 st transmission rate of 1-1.5, after 4-5 days of culture, the cells are fused at the ratio of 1: subculturing at a ratio of 6-8; and preparing a mesenchymal stem cell suspension: and (3) taking P4 generation cells in the culture and amplification of the primary mesenchymal stem cell to culture and amplify for 48-96 hours, after the cells are fused for no more than 80%, digesting the cells by using a third digestion solution, washing, suspending by using 5% human albumin, and adding physiological saline to obtain the required mesenchymal stem cell suspension. However, the patent does not modify DMEM, resulting in short shelf life and limited cell viability.
Disclosure of Invention
1. Problems to be solved
Aiming at the problems in the prior art, the invention provides the synovial membrane mesenchymal stem cell suspension and the preparation method thereof, which have the advantages of simple process and easy manufacture and are beneficial to improving the suspension stability of the synovial membrane mesenchymal stem cells.
2. Technical scheme
In order to solve the above problems, the present invention adopts the following technical solutions.
A preparation method of a synovial mesenchymal stem cell suspension comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
The synovial mesenchymal stem cell suspension is prepared by the preparation method of the synovial mesenchymal stem cell suspension.
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
the improved DMEM culture medium is innovatively introduced, the new additive zinc acetate plays a critical role, and the hyaluronic acid also plays a certain promoting role.
Drawings
FIG. 1 is a graph showing the results of laser confocal in example 1;
FIG. 2 is a graph showing the results of laser confocal in comparative example 1;
FIG. 3 is a graph showing the results of laser confocal in comparative example 2;
FIG. 4 is a graph showing the results of laser confocal in comparative example 3;
FIG. 5 is a graph showing the results of laser confocal in comparative example 4;
FIG. 6 is a graph showing the results of laser confocal in comparative example 5.
Detailed Description
The invention is further described with reference to specific examples.
It needs to be reminded that the synovial membrane mesenchymal stem cells researched by the application are mouse synovial membrane mesenchymal stem cells, are mainly used for researching the suspension stability of the mouse synovial membrane mesenchymal stem cells, and are combined with instruments such as a live-dead cell staining kit to detect the activity of the mouse synovial membrane mesenchymal stem cells.
Example 1
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 1
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 2
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
trypsin 1 mg.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 3
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 4
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
and recombining into fiber growth factor FGF-9100 ng.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 5
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing into a DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the cell fusion phenomenon occurs at 70% in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃, a DMEM medium and a 7d culture.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Example 6
The suspension prepared in example 1 and the suspensions prepared in comparative examples 1 to 5 were selected and tested as follows:
the suspension prepared above was stored at 4 ℃ for one month, and then the identification of stem cells in the suspension was observed using a live-dead cell staining kit (purchased from Eimei technologies, Inc., Wuhan) in combination with a laser confocal instrument.
As shown in fig. 1, the number of living cells is large, the number of dead cells is small, the morphology is normal, and the cell density is normal;
as shown in FIG. 2, the number of living cells is less than that of FIG. 1, the number of dead cells is more than that of FIG. 1, the morphology of part of the cells is distorted, the number of dead cells is more than that of living cells as a whole, and the cell density is less than that of FIG. 1;
as shown in fig. 3, the number of living cells is less than that of fig. 2, the number of dead cells is more than that of fig. 2, the number of dead cells is much more than that of living cells as a whole, the morphology of part of the cells is distorted, and the cell density is less than that of fig. 1;
as shown in fig. 4, the number of living cells was the smallest, the number of dead cells was greater than that of fig. 3, and the number of dead cells was 5 times as large as that of living cells as a whole, and the cell density was the smallest;
as shown in FIG. 5, the number of live cells is less than that of FIG. 3 and more than that of FIG. 4, the number of dead cells is 2 times that of dead cells as a whole, and the cell density is less than that of FIG. 4 and less than that of FIG. 3;
as shown in fig. 6, the number of live cells was between fig. 4 and fig. 5, the number of dead cells was equal to the number of live cells as a whole, and the cell density was between fig. 4 and fig. 5.
While the invention has been described in further detail in connection with specific embodiments thereof, it will be understood that the invention is not limited thereto, and that various other modifications and substitutions may be made by those skilled in the art without departing from the spirit of the invention, which should be considered to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. A method for preparing a suspension of synovial mesenchymal stem cells, which is characterized by comprising the following steps:
the method comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
2. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
3. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
4. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 3, wherein:
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
5. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
6. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃.
7. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
8. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the temperature for the treatment in step (3) was 10 ℃.
9. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the preservation temperature after the treatment in step (3) was 4 ℃.
10. A suspension of synovial mesenchymal stem cells, wherein:
the synovial mesenchymal stem cell suspension of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907400.0A CN113755433A (en) | 2021-08-09 | 2021-08-09 | Synovial mesenchymal stem cell suspension and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907400.0A CN113755433A (en) | 2021-08-09 | 2021-08-09 | Synovial mesenchymal stem cell suspension and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113755433A true CN113755433A (en) | 2021-12-07 |
Family
ID=78788762
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110907400.0A Pending CN113755433A (en) | 2021-08-09 | 2021-08-09 | Synovial mesenchymal stem cell suspension and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113755433A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115418348A (en) * | 2022-08-06 | 2022-12-02 | 北京永华燕芸生物科技有限公司 | Method for extracting and culturing synovial effusion mesenchymal stem cells |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
| US20150086514A1 (en) * | 2012-11-14 | 2015-03-26 | Zaimei JIA | Mesenchymal stem cell injection and preparation method thereof, and application thereof in preparing diabetes drug |
| CN105524878A (en) * | 2016-01-21 | 2016-04-27 | 深圳市第二人民医院 | Culture method for separating and inducting hUC-MSCs into cartilage cells |
| CN108456655A (en) * | 2018-02-23 | 2018-08-28 | 深圳至博生物科技有限公司 | Mescenchymal stem cell suspension and the preparation method and application thereof |
-
2021
- 2021-08-09 CN CN202110907400.0A patent/CN113755433A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
| US20150086514A1 (en) * | 2012-11-14 | 2015-03-26 | Zaimei JIA | Mesenchymal stem cell injection and preparation method thereof, and application thereof in preparing diabetes drug |
| CN105524878A (en) * | 2016-01-21 | 2016-04-27 | 深圳市第二人民医院 | Culture method for separating and inducting hUC-MSCs into cartilage cells |
| CN108456655A (en) * | 2018-02-23 | 2018-08-28 | 深圳至博生物科技有限公司 | Mescenchymal stem cell suspension and the preparation method and application thereof |
Non-Patent Citations (9)
| Title |
|---|
| HIDEYA YOSHIMURA等: "《Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle》", 《CELL TISSUE RES.》 * |
| JI HYUN KIM等: "《Enhanced proliferation and chondrogenic differentiation of human synovium-derived stem cells expanded with basic fibroblast growth factor》", 《TISSUE ENG PART A.》 * |
| MI-YOUNG MOON等: "《Zinc Promotes Adipose-Derived Mesenchymal Stem Cell Proliferation and Differentiation towards a Neuronal Fate》", 《STEM CELLS INTERNATIONAL》 * |
| SMITHA MATHEWS等: "《Glycosaminoglycans enhance osteoblast differentiation of bone marrow derived human mesenchymal stem cells》", 《JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE》 * |
| 庞雪芬: "《透明质酸对脂肪干细胞增殖和分化的影响》", 《中山大学硕士学位论文》 * |
| 张正政等: "《滑膜间充质干细胞分离培养、免疫表型鉴定及在混合淋巴反应体系中的抑制效应》", 《中国组织工程研究》 * |
| 张正政等: "滑膜间充质干细胞分离培养、免疫表型鉴定及在混合淋巴反应体系中的抑制效应", 《中国组织工程研究》 * |
| 符培亮等: "滑膜间充质干细胞分离纯化后的生物学特性", 《中国组织工程研究》 * |
| 龚忠诚等: "滑膜间充质干细胞软骨分化能力的实验研究", 《新疆医科大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115418348A (en) * | 2022-08-06 | 2022-12-02 | 北京永华燕芸生物科技有限公司 | Method for extracting and culturing synovial effusion mesenchymal stem cells |
| CN115418348B (en) * | 2022-08-06 | 2024-05-31 | 北京永华燕芸生物科技有限公司 | Extraction and culture method of synovial effusion mesenchymal stem cells |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114317443B (en) | Breast cancer organoid culture solution, and culture reagent combination and culture method thereof | |
| CN110938590B (en) | Mesenchymal stem cell serum-free medium and application thereof | |
| US20220325247A1 (en) | METHOD FOR PROMOTING GROWTH OF PARIETAL DECIDUAL BASALIS-MESENCHYMAL STEM CELLS (PDB-MSCs) | |
| CN110179826B (en) | Human umbilical cord mesenchymal stem cell source stem cell factor microvesicle preparation and preparation method thereof | |
| CN114292816A (en) | Lung cancer organoid culture solution, and culture reagent combination and culture method thereof | |
| CN113564111B (en) | Method for culturing umbilical cord-derived mesenchymal stem cells in low-oxygen mode | |
| CN110564681A (en) | Isolated culture and nerve directional differentiation method of deciduous tooth pulp stem cells | |
| CN113755433A (en) | Synovial mesenchymal stem cell suspension and preparation method thereof | |
| CN111424011A (en) | Three-dimensional culture method capable of maintaining cell morphology of umbilical cord mesenchymal stem cells | |
| CN112725270A (en) | Human-derived bone marrow mesenchymal stem cell induction culture medium and induction method | |
| CN106834223B (en) | Method for inducing differentiation of umbilical cord mesenchymal stem cells into chondrocytes | |
| CN112680408A (en) | Culture medium for inducing human mesenchymal stem cells to form cartilage differentiation and preparation method thereof | |
| CN112941017A (en) | Culture medium for inducing human mesenchymal stem cells to form fat and differentiate and preparation method thereof | |
| CN107164325B (en) | The preparation method and kit of the oligodendroglia in the source MSCs | |
| CN111534482B (en) | Culture medium for chondrogenic differentiation and chondrogenic differentiation of umbilical cord mesenchymal stem cells | |
| CN115067318A (en) | Dental pulp mesenchymal stem cell preservation solution and application thereof | |
| CN106957814B (en) | Culture medium for amniotic mesenchymal stem cells and method for culturing amniotic mesenchymal stem cells | |
| CN115835874A (en) | Method for producing synovial-derived mesenchymal stem cells and method for producing cell preparation for joint treatment | |
| CN105368772A (en) | Culture medium, application thereof and method for cultivating dental pulp stem cells | |
| CN110468097B (en) | Induction system and method for efficiently promoting chondrogenic differentiation of mesenchymal stem cells | |
| CN116536255B (en) | Culture medium and method for efficiently inducing muscle stem cells | |
| CN113717933B (en) | Application of FGF7 in preparation of stem cell expansion and phenotype maintaining reagent | |
| CN117187174B (en) | Muse cell culture medium and extraction method of fat Muse cells | |
| CN115369084A (en) | Adipose-derived mesenchymal stem cell culture medium and application thereof | |
| CN117568268A (en) | Culture method capable of maintaining tooth germ mesenchymal cells in vitro into tooth fate for long term and application thereof |
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: 20211207 |