Proliferation accelerator for human adipose-derived stem cells and application method thereof
A technology of human adipose stem cells and application methods, which is applied in the field of human adipose stem cell proliferation promoters, and can solve the problems of slow subculture proliferation, complicated methods, and small number of stem cells
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Embodiment 1
[0030] Embodiment 1: Common human adipose stem cell basal medium preparation:
[0031] Take 900 mL of DMEM / F12 cell culture medium (purchased from TAKARA BIO INC.) and add antibiotics: 90000 U of penicillin and 90000 U of streptomycin.
[0032] Adjust pH value: use mass fraction 5% NaHCO 3 Adjust the pH to 7.2.
[0033] Sterilization by filtration: Use a filter membrane with a pore size of 0.45 μm and 0.22 μm, the upper layer is 0.45 μm, and the lower layer is 0.22um to ensure the filtering effect.
[0034] Add calf serum: before use, add calf serum to make up to 1000mL (purchased from TAKARA BIO INC.).
Embodiment 2
[0035] Example 2: Preparation of a proliferation medium A containing human adipose stem cell proliferation promoter
[0036] If the predetermined volume is 1000mL, take 900mL of DMEM / F12 cell culture medium (TAKARA BIO INC.), add antibiotics: penicillin 90000U, streptomycin 90000U.
[0037] Adjust pH value: use mass fraction 5% NaHCO 3 Adjust the pH to 7.2.
[0038] Adding proliferation promoter: Weigh the components and amounts shown in Table 1 and add them to the prepared DMEM / F12 cell culture medium in sequence, shake or sonicate to help dissolve, and do not heat to help dissolve.
[0039] Table 1
[0040]
[0041] Then the concentration of each component shown in the above table in proliferation medium A is insulin 0.1 μg / mL, hydrocortisone 10 -10 mol / L, parathyroid hormone 2×10 -10mol / L, estradiol 0.2ng / mL, testosterone 0.8ng / mL, platelet-derived growth factor-AB 1ng / mL, epidermal growth factor 2ng / mL, basic fibroblast growth factor 2ng / mL, L-glutamine Amide 0.4mm...
Embodiment 3
[0044] Example 3: Preparation of a proliferation medium B containing human adipose stem cell proliferation promoter
[0045] The preparation method and predetermined amount are as in Example 2, but when weighing, the components in Table 1 can be used to make the concentrations in the proliferation medium B successively insulin 10 μg / mL, hydrocortisone 5×10 -9 mol / L, parathyroid hormone 7×10 -10 mol / L, estradiol 3.5ng / mL, testosterone 15ng / mL, platelet-derived growth factor-AB8ng / mL, epidermal growth factor 16ng / mL, basic fibroblast growth factor 20ng / mL, L-glutamine 4mmol / L, 2-mercaptoethanol 0.15mmol / L, leukemia inhibitory factor 7ng / mL, L-ascorbic acid 80μg / mL, biotin 45μg / mL, dexamethasone 5×10 -8 mol / L, IGF-I10ng / mL and SCF15ng / mL were weighed.
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