Serum-free adipose-derived stem cell culture medium and preparation method thereof
A technology of fat stem cells and culture medium, applied in the field of stem cells, can solve the problems of poor cell adhesion and unsatisfactory cell proliferation rate, and achieve the effect of simple preparation method, good cell differentiation potential, and promotion of growth and proliferation
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Embodiment 1
[0032] Example 1 Effect of media containing different concentrations of stem cell growth factors on cell proliferation
[0033] The inventor conducted a large number of tests to screen and obtain the serum-free fat stem cell culture medium formula provided by the present invention, and further investigated the influence of the concentration of stem cell growth factors. The P1 generation of ADSCs at 5000 / cm 2The density was inoculated in six-well plates, and cultured with serum-free adipose-derived stem cell culture medium containing different concentrations of stem cell growth factors (compared with medium 1, the difference was only the concentration of stem cell growth factors was different), and the cells were divided every 3 days. Digest with 0.25% trypsin, collect the cells and calculate the number, and use 5000 cells / cm 2 The density was re-inoculated and cultured until the P5 generation, and the cell proliferation curve was drawn according to the number of cells harve...
Embodiment 2
[0035] Example 2 Serum-free adipose stem cell culture medium
[0036] Serum-free fat stem cell culture medium, including IMDM basal medium, also includes the following components and their concentrations: recombinant human insulin 1mg / mL, recombinant transferrin 2.5μg / mL, vitamin C 16.1μg / mL, human serum albumin 1mg / mL, bFGF 10ng / mL, PDGF-bb 10ng / mL, EGF 10ng / mL, IGF-1 10ng / mL, TGF-β10ng / mL, fibronectin 10ng / mL, fetuin 1mg / mL, polytea Phenol 10μg / mL and stem cell growth factor 10ng / mL.
[0037] Preparation method: Add recombinant human insulin mother liquor, recombinant transferrin mother liquor, vitamin C mother liquor, human serum albumin, bFGF mother liquor, PDGF-bb mother liquor, EGF mother liquor, IGF-1 mother liquor to the IMDM basal medium according to the stated concentration , TGF-β mother solution, fibronectin mother solution, fetuin mother solution, tea polyphenol mother solution and stem cell growth factor mother solution, stir evenly, pass through the membrane ...
Embodiment 3
[0039] Example 3 Serum-free adipose stem cell culture medium
[0040] Serum-free fat stem cell culture medium, including IMDM basal medium, also includes the following components and their concentrations: recombinant human insulin 2mg / mL, recombinant transferrin 2μg / mL, vitamin C 20μg / mL, human serum albumin 1mg / mL mL, bFGF 15ng / mL, PDGF-bb 10ng / mL, EGF 15ng / mL, IGF-1 10ng / mL, TGF-β10ng / mL, fibronectin 15ng / mL, fetuin 2mg / mL, tea polyphenol 15μg / mL and stem cell growth factor 15ng / mL.
[0041] Preparation method: similar to the preparation method in Example 2.
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