Human adipose-derived stem cell serum-free basic medium
A technology of human adipose stem cells and basal medium, applied in the field of stem cell serum-free basal medium, can solve the problems of slow subculture proliferation, stem cells not suitable for direct application, and small number of stem cells, etc., and achieve the effect of accelerated growth
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Embodiment 1
[0026] Example 1: Preparation of conventional serum-containing basal medium
[0027] 1) Prepare a predetermined amount of 1000 mL, take 900 mL of high-glucose DMEM cell culture medium (manufacturer: Sigma, product number: D-5030), add 90,000 U of penicillin and 90,000 U of streptomycin.
[0028] 2) adjust the pH value: use 5% NaHCO 3 Adjust the pH to 7.2.
[0029] 3) Sterilization by filtration: Use one filter membrane of 0.45um and 0.22um each, the upper layer is 0.45um and the lower layer is 0.22um to ensure the filtering effect.
[0030] 4) Add calf serum: before use, add calf serum (manufactured by TAKARA BIOINC.) to a predetermined volume of 1000mL.
Embodiment 2
[0031] Example 2: Preparation of a serum-free basal medium A for human adipose stem cells
[0032] 1) Prepare a predetermined amount of 1000 mL, take 900 mL of high-glucose DMEM cell culture medium (manufacturer: Sigma, product number: D-5030), add 90,000 U of penicillin and 90,000 U of streptomycin.
[0033] 2) adjust pH value: use mass fraction 5% NaHCO 3 Adjust the pH to 7.2.
[0034] 3) Add serum replacement agent: Weigh all 10 reagents listed in Table 2 according to the following final concentration requirements:
[0035]
[0036] And sequentially added to the high-sugar DMEM cell culture medium prepared above, oscillated or ultrasonically aided in dissolving, without heating to assist in dissolving.
[0037] 4) Then add the high-glucose DMEM cell culture solution prepared above to the predetermined total solution volume of 1000mL.
Embodiment 3
[0038] Example 3: Preparation of a serum-free basal medium B for human adipose stem cells
[0039] Change all 10 reagents listed in Table 2 to weigh according to the following final concentration requirements:
[0040]
[0041] The predetermined amount and specific preparation method steps are as in Example 2.
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