Adipogenic differentiation

Similar to osteogenic differentiation, adipogenic differentiation and oil red O detection were performed using an adipogenic differentiation induction kit according to the manufacturer’s instructions. First, dexamethasone, rosiglitazone, 3-isobutyl-1-methylxanthine, insulin, penicillin/streptomycin, and FBS were added into the basal medium to form complete adipogenic differentiation induction medium A. Adipogenic differentiation maintenance medium B was formed by adding insulin, penicillin/streptomycin, and FBS to the basal medium. When hPDMSCs were 80% confluent, the cells were cultured with complete adipogenic induction medium A for 72 h, and then the medium was replaced with maintenance adipogenic induction medium B and further incubated for 24 h, which was one cycle. Three to five cycles were repeated. When obvious lipid droplets appeared in the cell, the cells were cultured only in adipogenic differentiation maintenance medium B, and new medium B was replaced every 2 days. When the lipid droplets were large enough, the culture ended. Then, the induced cells were fixed with paraformaldehyde, stained with oil red O, and imaged.