Two different hPSCs were used in this study: H9 (WA09; passage 35) was purchased from the WiCell Research Institute (Madison, WI, USA) and CHA-SCNT-PSC-18 (Korea Stem Cell Registry code hES12019001), which we previously established [33 (link),34 (link)]. H9 cells were maintained in the mTeSR medium (Stem Cell Technologies, BC, Canada). CHA-SCNT-PSC-18 was cultured on mitotically inactivated mouse embryonic fibroblasts (CF1; Jackson Laboratory, CA, USA) in Dulbecco’s modified Eagle medium (DMEM)/F12 (Nacalai Tesque, Kyoto, Japan) supplemented with 20% knockout serum replacement (KSR; Thermo Fisher Scientific), 0.1 mM beta-mercaptoethanol (Thermo Fisher Scientific, MA, US), 1% non-essential amino acids (NEAA; Thermo Fisher Scientific), and 4 ng/mL recombinant human FGF2 (Gibco). The hPSCs medium was changed daily, and cells were passaged every 4–5 days. The hPSCs were maintained on Matrigel (BD Biosciences, San Jose, CA, USA)-coated plates in mTeSR and passaged when reaching 70–80% confluency. Cells before passage 45 were used to generate hSkMOs.
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