Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes

• DOI: 10.1016/j.stemcr.2017.08.025
• Publication Date: 2017-10-11T10:16:32Z
• Authors: Shugo Tohyama; Jun Fujita; Chihana Fujita; Miho Yamaguchi; Sayaka Kanaami; Rei Ohno; Kazuho Sakamoto; Masami Kodama; Junko Kurokawa; Hideaki Kanazawa; Tomohisa Seki; Yoshikazu Kishino; Marina Okada; Kazuaki Nakajima; Sho Tanosaki; Shota Someya; Akinori Hirano; Shinji Kawaguchi; Eiji Kobayashi; Keiichi Fukuda
• Keywords: 0301 basic medicine; Medicine (General); QH301-705.5; Induced Pluripotent Stem Cells; Primary Cell Culture; Cell Differentiation; Culture Media; 03 medical and health sciences; R5-920; Report; Humans; Myocytes, Cardiac; Biology (General); Cells, Cultured; Cell Proliferation

Cardiac regenerative therapies utilizing human induced pluripotent stem cells (hiPSCs) are hampered by ineffective large-scale culture. hiPSCs were cultured in multilayer culture plates (CPs) with active gas ventilation (AGV), resulting stable proliferation and pluripotency. Seeding of 1 × 106 per layer yielded 7.2 108 4-layer CPs 1.7 109 10-layer sequentially differentiated into cardiomyocytes (CMs) a two-dimensional (2D) differentiation protocol. The efficiency cardiac using AGV was 66%–87%. Approximately 6.2–7.0 (4-layer) 1.5–2.8 (10-layer) obtained AGV. After metabolic purification glucose- glutamine-depleted lactate-supplemented media, massive amount purified CMs prepared. Here, we present scalable 2D system for hiPSC-derived CMs, which will facilitate clinical applications severe heart failure the near future.