CN110343659A - A kind of mescenchymal stem cell complete medium composition - Google Patents
A kind of mescenchymal stem cell complete medium composition Download PDFInfo
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- CN110343659A CN110343659A CN201810306206.5A CN201810306206A CN110343659A CN 110343659 A CN110343659 A CN 110343659A CN 201810306206 A CN201810306206 A CN 201810306206A CN 110343659 A CN110343659 A CN 110343659A
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- stem cell
- mescenchymal stem
- antibiotic
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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
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
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- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/44—Thiols, e.g. mercaptoethanol
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- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/80—Undefined extracts from animals
- C12N2500/84—Undefined extracts from animals from mammals
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
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- 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
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
Abstract
The invention discloses a kind of mescenchymal stem cell complete medium compositions, using DMEM culture medium as solvent, and contain following concentration component: 2~15wt% of fetal calf serum, 20~100nM/L of beta -mercaptoethanol, 5~50ng/mL of FGF, EGF5~50ng/mL, 1 × NEAA and 1 × antibiotic.Culture medium of the invention is based primarily upon mescenchymal stem cell and needs to prepare to nutrition owner, can maintain the form of mescenchymal stem cell, and keep the Multidirectional Differentiation of mescenchymal stem cell, furthermore the preparation method of culture medium of the invention is simple, low in cost.
Description
Technical field
The invention belongs to technical field of cell culture, and in particular to a kind of mescenchymal stem cell complete medium composition.
Background technique
Mescenchymal stem cell culture medium be carry out mescenchymal stem cell therapeutic test basis, mesenchymal cell state it is good
It is bad, be directly related to research as a result, mescenchymal stem cell in good condition is the basis of stem cell repair function.Mesenchyma is dry
Cell has Multidirectional Differentiation, can be divided into lipoblast, osteoblast and chondroblast under certain condition.It is undesirable
Condition of culture can lead to stem cell and be divided into other cells, while can also cause the aging of mescenchymal stem cell, to lose it
Repair function.With the development of stem cell, the research that stem cell is applied to treatment heart disease is also more and more, and stem cell is used for
The research of tissue repair becomes the hot spot of regenerative medicine, repairs the research such as myocardial infarction, renal failure treatment, neural restoration with mesenchyma
Based on.However, it is inconsistent often to will appear result in stem cell incubation, it can not polyisomenism.This phenomenon and cell
Close relation is cultivated, its activity of the stem cell of different conditions and repair are also different, so between Selective agar medium becomes
The pith of mesenchymal stem cells repairing research is directly related to the splendid attire state of cell.
Summary of the invention
It is an object of the invention to overcome prior art defect, a kind of mescenchymal stem cell complete medium combination is provided
Object.
Technical scheme is as follows:
A kind of mescenchymal stem cell complete medium composition, using DMEM culture medium as solvent, and containing following concentration
Component: 2~15wt% of fetal calf serum, 20~100nM/L of beta -mercaptoethanol, 5~50ng/mL of FGF, 5~50ng/mL of EGF, 1
× NEAA and 1 × antibiotic；
The preparation method of above-mentioned mescenchymal stem cell complete medium composition are as follows: by fetal calf serum, beta -mercaptoethanol,
FGF, EGF, 1 × NEAA and 1 × antibiotic be dissolved in DEME culture medium, after filtration sterilization.
In a preferred embodiment of the invention, using DMEM culture medium as solvent, and contain the component of following concentration:
8~12wt% of fetal calf serum, 40~60nM/L of beta -mercaptoethanol, 5~15ng/mL of FGF, 5~15ng/mL of EGF, 1 ×
NEAA and 1 × antibiotic.
It is further preferred that using DMEM culture medium as solvent, and the component containing following concentration: fetal calf serum 10wt%,
Beta -mercaptoethanol 50nM/L, FGF 10ng/mL, EGF 10ng/mL, 1 × NEAA and 1 × antibiotic.
In a preferred embodiment of the invention, the antibiotic is antibiotic-antimycotic.
The beneficial effects of the present invention are:
1, culture medium of the invention is based primarily upon mescenchymal stem cell and needs to prepare to nutrition owner, fills between can maintaining
The form of matter stem cell, and the Multidirectional Differentiation of mescenchymal stem cell is kept, the furthermore preparation method letter of culture medium of the invention
It is single, it is low in cost.
2, preparation method of the invention is simple, low in cost.
Detailed description of the invention
Fig. 1 is the culture photo of the human umbilical cord mesenchymal stem cells in the embodiment of the present invention 1.
Fig. 2 is the culture photo of Rat Mesenchymal Stem Cells in the embodiment of the present invention 2, wherein A: rat MSC morphology is seen
It examines；B: rat MSC breaks up to chondroblast；C: rat MSC to osteoblast differentiation；D: rat MSC to Adipocyte Differentiation.
Fig. 3 is the culture photo of the mouse mesenchymal cell in the embodiment of the present invention 3.
Fig. 4 is the Rat Mesenchymal Stem Cells streaming figure in the embodiment of the present invention 2.
Specific embodiment
Technical solution of the present invention is further explained and described below by way of specific embodiment combination attached drawing.
Embodiment 1
Mescenchymal stem cell complete medium formula: using DMEM culture medium as solvent, and contain the component of following concentration: tire
Cow's serum 10wt%, beta -mercaptoethanol 50nM/L, FGF 10ng/mL, EGF 10ng/mL, 1 × NEAA and 1 ×
antibiotic-antimycotic。
Its preparation method is as follows: by fetal calf serum, beta -mercaptoethanol, FGF, EGF, 1 × NEAA and 1 ×
Antibiotic-antimycotic is dissolved in DEME culture medium, after filtration sterilization.
Human umbilical cord mesenchymal stem cells are separated, with above-mentioned culture medium, 37 DEG C, are cultivated under 5% carbon dioxide conditions,
It can be seen that human mesenchymal stem cell is in shuttle shape triangle, and adherent growth is good (see Fig. 1)
Embodiment 2
Mescenchymal stem cell complete medium formula: using DMEM culture medium as solvent, and contain the component of following concentration: tire
Cow's serum 10wt%, beta -mercaptoethanol 50nM/L, FGF 10ng/mL, EGF 10ng/mL, 1 × NEAA and 1 ×
antibiotic-antimycotic。
Its preparation method is as follows: by fetal calf serum, beta -mercaptoethanol, FGF, EGF, 1 × NEAA and 1 ×
Antibiotic-antimycotic is dissolved in DEME culture medium, after filtration sterilization.
Rat bone marrow mesenchymal stem cells are extracted, with above-mentioned culture medium, 37 DEG C, are trained under 5% carbon dioxide conditions
Support, it can be seen that Rat Mesenchymal Stem Cells be in shuttle shape triangle, it is in good condition, and can be divided into lipoblast, osteoblast,
Chondroblast (see Fig. 2), and carry out the identification of streaming surface (see Fig. 4).
Embodiment 3
Mescenchymal stem cell complete medium formula: using DMEM culture medium as solvent, and contain the component of following concentration: tire
Cow's serum 10wt%, beta -mercaptoethanol 50nM/L, FGF 10ng/mL, EGF 10ng/mL, 1 × NEAA and 1 ×
antibiotic-antimycotic。
Its preparation method is as follows: by fetal calf serum, beta -mercaptoethanol, FGF, EGF, 1 × NEAA and 1 ×
Antibiotic-antimycotic is dissolved in DEME culture medium, after filtration sterilization
.Marrow Mesenchymal Stem Cells are extracted, with above-mentioned culture medium, 37 DEG C, are trained under 5% carbon dioxide conditions
It supports, it can be seen that mouse mesenchymal cell is in shuttle shape triangle, (see Fig. 3) in good condition.
Those of ordinary skill in the art still are able to it is found that when technical solution of the present invention changes in following ranges
To same as the previously described embodiments or similar technical effect, protection scope of the present invention is still fallen within:
A kind of mescenchymal stem cell complete medium composition, using DMEM culture medium as solvent, and containing following concentration
Component: 2~15wt% of fetal calf serum, 20~100nM/L of beta -mercaptoethanol, 5~50ng/mL of FGF, 5~50ng/mL of EGF, 1
× NEAA and 1 × antibiotic；Preferably, using DMEM culture medium as solvent, and contain the component of following concentration: fetal calf serum
8~12wt%, 40~60nM/L of beta -mercaptoethanol, 5~15ng/mL of FGF, 5~15ng/mL of EGF, 1 × NEAA and 1 ×
Antibiotic.
The foregoing is only a preferred embodiment of the present invention, the range that the present invention that therefore, it cannot be limited according to is implemented, i.e.,
Equivalent changes and modifications made in accordance with the scope of the invention and the contents of the specification should still be within the scope of the present invention.
Claims (4)
1. a kind of mescenchymal stem cell complete medium composition, it is characterised in that: using DMEM culture medium as solvent, and containing such as
The component of lower concentration: 2~15wt% of fetal calf serum, 20~100nM/L of beta -mercaptoethanol, 5~50ng/mL of FGF, EGF 5~
50ng/mL, 1 × NEAA and 1 × antibiotic；
The preparation method of above-mentioned mescenchymal stem cell complete medium composition are as follows: by fetal calf serum, beta -mercaptoethanol, FGF,
EGF, 1 × NEAA and 1 × antibiotic be dissolved in DEME culture medium, after filtration sterilization.
2. a kind of mescenchymal stem cell complete medium composition as described in claim 1, it is characterised in that: cultivated with DMEM
Base is solvent, and the component containing following concentration: 8~12wt% of fetal calf serum, 40~60nM/L of beta -mercaptoethanol, FGF 5~
5~15ng/mL of 15ng/mL, EGF, 1 × NEAA and 1 × antibiotic.
3. a kind of mescenchymal stem cell complete medium composition as claimed in claim 2, it is characterised in that: cultivated with DMEM
Base is solvent, and contains the component of following concentration: fetal calf serum 10wt%, beta -mercaptoethanol 50nM/L, FGF 10ng/mL, EGF
10ng/mL, 1 × NEAA and 1 × antibiotic.
4. a kind of mescenchymal stem cell complete medium composition as described in claim 1, it is characterised in that: the antibiotic
For antibiotic-antimycotic.
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| US20140248653A1 (en) * | 2010-06-21 | 2014-09-04 | Centre D'etude Des Cellules Souches | Method For Selecting Mevalonate Synthesis Modulators Using Cells Derived From Human Pluripotent Cells |
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