193. Establishment of an Optimization Procedure for Large-Scale Production of Patient-Specific Cardiomyocytes from hiPSCs

Abstract

Cardiovascular diseases are a leading cause of morbidity and mortality worldwide. Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), with their ability of indefinite self-renewal and capability to differentiate into all three germ layers, provide a powerful tool in developmental biology, for drug screening, disease modeling, and potentially cell replacement therapy for diseases, such as myocardial infarction and heart failure. Unlike hESCs, hiPSCs are generated from reprogrammed somatic cells without using of human embryos and thus hold a great promise for cell-based therapy as they avoid ethical and immune rejection issues. However, due to scalability limitation of current hiPSC adherent culture platform and potential variation of culture and differentiation conditions among hiPSC lines, production of hiPSC-derived cells to a scale sufficient for therapeutic application in a timely manner has been challenging. Previously we have developed a scalable, defined, and cGMP-compliant suspension culture system for hPSC expansion and cardiac differentiation using spinner flasks by optimizing critical culture parameters we identified. In the present study, we tested the feasibility of applying our suspension culture platform and optimization strategy to establish a process for scale-up production of cardiomyocytes (CM) from individual hiPSC lines in a timely manner. We optimized cell seeding density, agitation rate, concentration of differentiation inducer, and timing for differentiation induction to determine an optimal condition for large-scale CM production. The process from suspension adaptation, differentiation optimization to CM production was accomplished in 3 months. With optimized culture and differentiation conditions, we showed that the hiPSC suspension culture can be serially passaged and expanded, and maintained normal karyotype. At a scale of 500mL suspension culture, 4-5×108 cells with 70-90% CM purity were manufactured. In summary, we demonstrated the feasibility of applying the suspension culture platform and optimization strategy we developed to establish a production process for individual hiPSC line to manufacture CM in a large scale in a timely manner.