iPS cells were cultured on Matrigel™ Matrix (Corning Life Sciences; catalog# 354234)-coated plates in mTeSR™ medium (Stem Cell Technologies; catalog# 85851). Matrigel was used at a 1:100 dilution in Minimum Essential Medium (MEM; Gibco, catalog# 51200038). The medium was refreshed daily and the cultures were passaged as single cells every 4–6 days. Briefly, for passaging and differentiation, iPS cells were washed with MEM and dissociated using Accutase™ (Stem Cell Technologies; catalog# AT-104). Cells were resuspended in mTeSR™ with 5 μM ROCK inhibitor (Stemolecule™ Y27632, Stemgent). iPS cells were re-plated dropwise to a Matrigel-coated dish containing mTeSR™ and 5 μM ROCK inhibitor at a density of 20,000 cells/cm2 for maintenance cultures and 50,000 cells/cm2 for differentiation.
