Differentiation and Induction of Mesenchymal Cells

EpiSCs differentiation was induced with SMEM (Gibco, Rockville, MD, USA) containing 0.05mM Ca²⁺, 10% FBS, 1% ampicilin/streptomycin in non-coated plates as described in [3 (link)]. SKPs differentiation was induced with DMEM:F12 at a ratio of 3:1 (Gibco, Rockville, MD), 15% fetal bovine serum (FBS), in poly-L-lysine coated plates (catalog number # P7280, Sigma, USA) as described in [12 (link),13 (link)]. The mesenchymal cells were induced to adipogenesis with DMEM-high glucose (Gibco, Rockville, MD), 10%FBS, 1uM dexamethasone (catalog number # D4902-25MG, Sigma, USA), 0.5mM IBMX (catalog number # I7018-250MG, Sigma, USA), 10ug/ul Insulin (catalog number # I1882-100MG, Invitrogen, USA), 100uM indomethacin (catalog number # I7378-5G, Sigma, USA). Chondrogenesis: DMEM-F12 (Gibco, Rockville, MD) 3:1, 0.5ug/ml Insulin (catalog number # I1882-100MG, Invitrogen, USA), 50uM ascorbic acid (catalog number # A4544-25G, Sigma, USA), 10ng/mL TGFβ-1 (catalog number # 7666-MB-005, R&D Systems, USA) and 1mM sodium piruvate (catalog number # 11360–070, Sigma, USA). Osteogenesis: αMEM (Gibco, Rockville, MD), 10% FBS, 0.1uM dexamethasone (catalog number # D4902-25MG, Sigma, USA), 2mM ascorbic acid (catalog number # A4544-25G, Sigma, USA), and 10mM Glycerol 2-Phosphate (catalog number # G9422-10G, Sigma, USA) as described in [19 (link)].