To induce differentiation of hESCs, cells cultured in mTeSR1 medium were dissociated by using Accutase (Stem Cell Technologies, Vancouver, BC, Canada) and washed three times with plain DMEM/F12 medium. Harvested cells were resuspended in hESC growth medium without bFGF supplement and transferred to ultra-low attachment plates to form embryoid bodies (Corning, Inc., Corning, NY). Aggregated embryoid bodies were kept for designated time period up to 10 days with daily media change.
