iPSC Maintenance Protocol for Diverse Cell Lines

Cell lines used were IMR90-4 iPSCs [32 (link)], CD12 iPSCs [33 (link)], CC3 iPSCs [34 (link)], and SM14 iPSCs [35 (link), 36 (link)]. IMR90-4 and CC3 lines are female. CD12 and SM14 lines are male. All cell lines were thawed directly into E8 medium containing 10 μM Y-27632 (Tocris). 24 h after thaw, cells were changed to E8 medium without Y-27632. E8 medium was changed every day thereafter. All iPSCs were maintained on Matrigel (Corning). Once 70% confluent, iPSCs were passaged by washing cells once with Versene solution (Thermo Fisher Scientific), incubating cells with Versene solution for 5 min at 37 °C, collecting the cells in fresh E8 medium, and distributing the cells at desired ratios to Matrigel-coated plates.