Culturing Stem Cells and Vascular Cells

H9 human ESCs purchased from WiCell Research institute were cultured on mitomycin C-inactivated mouse embryonic fibroblasts (MEFs) or matrigel (BD Biosciences) (Liu et al., 2012 (link)). Human VECs were cultured in EGM-2 (Lonza) medium supplemented with 50 ng/mL VEGF (HumanZyme), 10 μmol/L SB431542 (Selleck) and 20 ng/mL FGF2 (Joint Protein Central, JPC). Human VSMCs were cultured in N2B27 medium (1:1 (v/v) of DMEM/F12 medium (Gibco) and Neurobasal medium (Gibco) supplemented with 2% B27 (Gibco), 1% N2 (Gibco), 55 μmol/L β-mercaptoethanol (Gibco) and 1% penicillin/streptomycin (Gibco)) supplemented with 10 ng/mL PDGF-BB (Peprotech). Human MSCs were cultured in 90% α-MEM medium (Gibco) supplemented with 10% fetal bovine serum (Ausbian, VS500T), 1% penicillin/streptomycin (Gibco), and 1 ng/mL FGF2 (JPC).