Cell culture and differentiation

The C3H10T1/2 cell line was obtained from the Center for Type Culture Collection of China. Cells were cultured in complete medium containing Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Gaithersburg, MD, USA) with 10% fetal bovine serum (FBS; Gibco) and penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA), and used for the in vitro experiments. To induce osteogenic differentiation, cells were cultured in osteogenic media containing 100 nM dexamethasone (Sigma-Aldrich, St. Louis, MO, USA), 50 μg/ml ascorbic acid (Sigma-Aldrich), and 10 mM β-glycerophosphate (Sigma-Aldrich). To induce adipogenic differentiation, cells were cultured in adipogenic medium containing 10 μg/ml insulin (Sigma-Aldrich), 1 μM dexamethasone (Sigma-Aldrich), 0.5 mM 3-isobutyl-1-methylxanthine (Sigma-Aldrich), and 1 μM rosiglitazone (Sigma-Aldrich). All cell-based in vitro experiments were repeated three times.