Multilineage Differentiation of Stem Cells

Cells were planked into plates containing basal complete medium and cultured to 80%–90% confluency. The basal medium was then replaced with the corresponding differentiation-inducing mixture. The medium was replaced with a DMEM with low glucose (Hyclone, New York, United States) complete medium containing 10 nM dexamethasone (Sigma, United States), 50 mg/mL ascorbic acid 2-phosphate (Sigma, United States), and 10 mM β-glycerophosphate (Sigma, United States) to induce osteogenic differentiation. Adipogenic differentiation induction medium contained 100 nM dexamethasone (Sigma, United States), 50 μg/mL ascorbic acid 3-phosphate (Sigma, United States), and 50 μg/mL indomethacin (Sigma, United States). Chondrogenic differentiation was induced using a chondrogenic differentiation medium (Lonza, Basel, Switzerland) supplemented with recombinant TGFb3 protein (R&D Systems, Minneapolis, MN, United States) (Li et al., 2020 (link)).