“…The cells were cultured in a medium that contained osteogenic (10 À7 mol l À1 dexamethasone, 10 mM b-glycerophosphate and 50 mg l À1 ascorbate-phosphate), adipogenic (10 À6 mol l À1 dexamethasone and 10 mg l À1 Insulin) or neurogenic (DMEM/F12 with 0.5% B27, 1% fetal bovine serum, 5% horse serum, 0.5 mM retinoic acid, 20 ng per ml of epidermal growth factor, 50 ng per ml of basic fibroblast growth factor, 50 ng per ml of nerve growth factor) materials. [35][36][37] Regents were mostly ordered from Sigma Aldrich (St Louis, MO, USA), except B27 (Invitrogen), epidermal growth factor and nerve growth factor (Peprotech). Two weeks later, osteogenic differentiation was assessed by Alizarin red staining, and intracellular lipid accumulation was visualized using Oil-Red-O staining and inverted phase contrast microscopy.…”