The primary goal of this project is to develop tissue-engineered pulsatile conduits TEPCs to treat single ventricle congenital heart defect using cardiomyocytes CMs derived from human induced pluripotent stem cells hiPSCs. The PIs group has successfully generated unlimited numbers of functional hiPSC-CMs based on robust cardiac differentiation and lactate-based metabolic selection. Engineered heart tissues EHTs made by seeding hiPSC-CMs and primary human cardiac fibroblasts into decellularized porcine myocardium matrix has produced robust contractility. The PI has also successfully scaled up EHTs 15x14.5mm by seeding seven million hiPSC-CMs and three million human cardiac fibroblasts. Additionally, the PIs group has developed novel fibroblast-derived biological glue to wrap EHTs onto the decellularized umbilical artery scaffold effectively to generate TEPCs. Moreover, the PI has established an efficacious bioreactor approach with which TEPCs can be cultured under conditions of biomechanical stretch that enhance the development of contractility and pressure generation of TEPCs. With robust hiPSC-CM derivation, efficient EHT generation and the novel bioreactors that provide biomechanical training to mimic native heart tissue formation, the PI is poised to develop efficacious TEPCs and then evaluate the therapeutic efficacy by implanting TEPCs as the venous interposition grafts in rats in the coming research period.