Final optimized media formulations for each of the three steps of cardiac differentiation.

The basal medium RPMI 1640 was found to be most successful for cardiac induction during phase 2 and phase 3 and therefore was maintained for phase 4. The addition of the thiol 1-thioglycerol was found to be essential for hEB formation and survival. High concentrations of PVA enhanced hiPSC hEB formation. Insulin was essential for hEB formation. The combination of BMP4 and FGF2 was optimal for mesoderm specification whilst FBS or human serum was essential in phase 3 for