Adipogenic differentiation basal medium a

Passage 3 hADSCs were counted and seeded at a density of 10⁵ per well in a six-well plate. When the cells reached 100 % confluence, Adipogenic Differentiation Basal Medium A supplemented with 10 % FBS, 1 % penicillin-streptomycin, dexamethasone, isobutylmethylxanthine (IBMX), insulin, glutamine, and Rosiglitazone (Cyagen Biosciences) was added to four wells and complete culture medium (DMEM-LG) was added to other two wells as the negative controls. Three days later the medium was changed to hADSC Adipogenic Differentiation Basal Medium B, which contains 10 % FBS, dexamethasone, insulin, and glutamine (Cyagen Biosciences). The medium was changed 24 h later to medium A. Medium A and B were alternated 3–5 times (12–20 days), and then medium B was maintained for 4–7 days until the lipid droplets were big and round enough. During maintained culture, medium B was changed every 2–3 days with fresh medium B. Oil red O staining assessed the differentiation potential of adipogenesis formation of intracellular lipid droplets.