For EC and pericyte differentiation, iPSCs were detached with Versene (0.48 mM EDTA solution; Life Technologies) and plated onto Matrigel (growth factor reduced, 1:60 dilution; Life Technologies)-coated plates at a density of approximately 70,000 cells/cm2 in mouse embryonic fibroblast conditioned medium (MEF-CM; DMEM [Nacalai Tesque]) containing 10% fetal calf serum, 2 mM L-glutamine, and 1% non-essential amino acids (NEAA) (Life Technologies) with 4 ng/mL bFGF (Wako Pure Chemicals Industries) for 2 days before induction. Cells were covered with Matrigel (1:60 dilution) 1 day before induction. MEF-CM was replaced with RPMI + B27 medium (RPMI-1640, 2 mM L-glutamine, B27 supplement without insulin) supplemented with 100 ng/mL of activin A (R&D Systems) for 1 day, followed by 10 ng/mL bone morphogenetic protein 4 (R&D), 10 ng/mL bFGF, and Matrigel (1:60 dilution) for 3 days without culture medium change. After differentiation at 4 days, the culture medium was replaced with RPMI + B27 medium with 100 ng/mL VEGF165 (Wako) and 1 mM 8-bromoadenosine-3′,5′-cyclic monophosphate sodium salt (8-bromo-cAMP) (Nacalai Tesque) for 1 day. Primitive mesoderm cells were purified with autoMACS (Miltenyi Biotec) using anti-KDR
