Application of melatonin in preparation of medicine for preventing premature senility of mesenchymal stem cells
A technology of stem cells and melatonin, applied in the field of medical applications, can solve problems such as the impact that has not yet been elucidated, and achieve the effects of increasing cell proliferation potential, enhancing osteogenic differentiation ability, and reducing the positive rate of staining
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[0032] 1. The effect of melatonin on the proliferation ability, positive rate of SA-β-Gal staining and cell cycle of premature aging mesenchymal stem cells.
[0033] Human BM-MSCs were stored at 37°C, 5% CO 2 Cultivated at 175cm in the incubator 2 In the culture bottle (the volume ratio of each component of the medium is 89% α-MEM, 10% fetal bovine serum, 1% penicillin, streptomycin), the medium is changed every three days. When the cell density is ≥90%, digest the cells with 0.25% trypsin-EDTA at 3000 / cm 2 They were inoculated in well plates of three sizes (96-well plates for proliferation experiments, 12-well plates for SA-β-gal staining, and 6-well plates for cell cycle detection) for the next experiment. When the cell density reaches 50%, add 200 μM HO 2 o 2 (diluted in growth medium), incubate for 2 h. After 2 hours, wash twice with serum-free α-MEM medium, and then add melatonin with different concentration gradients (10nM, 1uM, 100uM) in complete medium to contin...
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