Schematic diagram of mitochondrial metabolic and redox signaling pathways during cardiomyocyte differentiation. A, During differentiation from Flk1+ MPCs to cardiomyocytes, cellular metabolism is changed from glycolysis to oxidative phosphorylation. CsA binds to cycophilin D which is a mitochondrial matrix protein that inhibits mPTP. Inhibition of mPTP activates the mitochondrial oxidative metabolism and promotes cardiomyocyte differentiation from Flk1+ MPCs. Addition of antioxidant with CsA profoundly augments cardiomyocyte differentiation. B, Estimation and comparison of the absolute number of cardiomyocytes generated from one ESC differentiated under control vehicle, CsA and CsA with antioxidants treatment. CsA indicates cyclosporin A; ESC, embryonic stem cell; IMM, inner mitochondrial membrane; MPCs, mesodermal precursor cells; mPTP, mitochondrial permeability transition pore; OMM, outer mitochondrial membrane; PSC, pluripotent stem cell; ROS, reactive oxygen species.