in hES+bFGF media. After 3 days, bFGF was withdrawn from the media and iPSCs were allowed to differentiate. Media was changed every 2 days. After 7 days, cultures were switched to NPC induction media and changed every other day until neural rosettes appeared. Neural rosettes were manually picked and transferred to poly-ornithine/laminin coated plates and cultured in NPC media. Cultures were fed every other day until confluent. Cells were passaged using TryplE at room temperature and plated at a density of 8×104 cells/cm2.
