For spontaneous embryoid body (EB) formation, hPSCs were manually passaged to low attachment plates in hESC medium without bFGF for 7 days, changing media every other day. On Day 8, EBs were transferred onto gelatin coated coverslips and cultured in the same medium for 7 more days. Directed differentiation was performed as previously described (Harness et al., 2011; Nistor et al., 2005).
