Pericytes were seeded in 24-well plates at 1 × 104 cells/well. Cells were cultured in Minimum Essential Medium Eagle (α-MEM, Wako) containing 10% FBS, 50 μM ascorbic acid 2-phosphate (Sigma-Aldrich), 5 mM disodium β-glycerophosphate (Sigma-Aldrich), 100 nM dexamethasone (Sigma-Aldrich) and 0.3 mg/mL recombinant human BMP-2 (R&D Systems, Minneapolis, MN, USA) for 6–9 days. The culture medium was changed every 3 days, and ALP activity and mineralization potential were evaluated by an ALP assay and von Kossa staining, respectively, as previously described [38 (link)].
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