Tesr e8 medium

H9 hESCs (WiCell Research Institute, Madison, WI, USA, Passages 45‐60) were cultured on Matrigel‐coated culture dishes (BD Biosciences, San Jose, CA, USA) with TeSR™‐E8™ medium (STEMcell Technologies, Vancouver, BC, Canada). Cells were passaged every 5 to 7 days by treatment with 0.5 mmol/L EDTA (Invitrogen, Waltham, MA, USA) for 3 minutes at 37°C and were transferred at a 1:20 ratio onto Matrigel‐coated plates with TeSR™‐E8™ medium containing 10 μmol/L Y‐27632 (Sigma‐Aldrich, St. Louis, MO, USA). Two days after passage, the medium was changed daily with fresh TeSR™‐E8™ medium without Y‐27632.

H9 human ESCs (WiCell Research Institute, Madison, WI, USA) were routinely maintained on Matrigel-coated culture dishes (BD Biosciences, San Jose, CA, USA) in TeSR™-E8™ medium (STEMCELL Technologies, Vancouver, BC, Canada). For passaging, ESCs were incubated with 0.5 mM EDTA (Thermo Fisher Scientific, Waltham, MA, USA) in a 37 °C CO₂ incubator for 3 min and then were split in the ratio of 1:20 onto Matrigel-coated dishes with TeSR™-E8™ medium containing 10 μM Y-27632 (Sigma-Aldrich, St. Louis, MO, USA). TeSR™-E8™ medium without Y-27632 was changed daily from two days after passage. Experiments using hESCs were approved by the Institutional Review Board of CHA University (IRB No. 1044308-201603-LR-004-09).