Metabolic Selection System for Large Numbers of Human iPSC-Derived Cardiomyocytes Shugo Tohyama1,2, Jun Fujita1, Chihana Fujita1, Miho Yamaguchi1, Rei Ohno1, Keiichi Fukuda1 1Department of Cardiology, Keio University School of Medicine 2Department of Organ Fabrication, Keio University School of Medicine
Heart regenerative therapy using human induced pluripotent stem cells (hiPSCs) is a potentially promising strategy for patients with heart disease, whereas the inability to eliminate residual hiPSCs and generate a massive amount of pure cardiomyocytes has been a barrier for realizing this potential. Recently, we established a novel method for purifying hiPSC-derived cardiomyocytes by focusing on glucose, glutamine and lactate metabolism in hiPSCs and differentiated cardiomyocytes (Tohyama S, Cell Stem Cell 2013, Cell Metabolism 2016). However, there are no efficient two-dimensional culture systems to obtain a large amount of pure cardiomyocytes. Therefore, we developed an advanced two-dimensional culture system using multilayer culture plates with active gas ventilation (AGV) that yielded a large number of hiPSCs and pure cardiomyocytes. Approximately 6 × 10E8 cells (four-layer) and 1.5 × 10E9 cells (10-layer) were obtained with AGV. After metabolic purification with glucose- and glutamine-depleted and lactate-supplemented media, a large amount of pure cardiomyocytes was finally prepared. This advanced cell culture system will facilitate the clinical application of hiPSC-derived cardiomyocytes.
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