@article{aa0ff83cbc0b49688fe8c76fb0d631ee,
title = "Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes",
abstract = "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 in stable proliferation and pluripotency. Seeding of 1 × 106 hiPSCs per layer yielded 7.2 × 108 hiPSCs in 4-layer CPs and 1.7 × 109 hiPSCs in 10-layer CPs with pluripotency. hiPSCs were sequentially differentiated into cardiomyocytes (CMs) in a two-dimensional (2D) differentiation protocol. The efficiency of cardiac differentiation using 10-layer CPs with AGV was 66%–87%. Approximately 6.2–7.0 × 108 cells (4-layer) and 1.5–2.8 × 109 cells (10-layer) were obtained with AGV. After metabolic purification with glucose- and glutamine-depleted and lactate-supplemented media, a massive amount of purified CMs was prepared. Here, we present a scalable 2D culture system using multilayer CPs with AGV for hiPSC-derived CMs, which will facilitate clinical applications for severe heart failure in the near future. In this article, Fujita and colleagues show that human induced pluripotent stem cells (hiPSCs) and hiPSC-derived cardiomyocytes (hiPSC-CMs) are stably and efficiently produced using multilayer culture plates with an active gas ventilation system because of stable CO2 and O2 concentration in culture media. Large numbers of obtained hiPSC-CMs are metabolically purified under glucose- and glutamine-depleted and lactate-supplemented conditions.",
keywords = "2D culture, cardiomyocytes, iPSCs, large scale, purification, regenerative medicine",
author = "Shugo Tohyama and Jun Fujita and Chihana Fujita and Miho Yamaguchi and Sayaka Kanaami and Rei Ohno and Kazuho Sakamoto and Masami Kodama and Junko Kurokawa and Hideaki Kanazawa and Tomohisa Seki and Yoshikazu Kishino and Marina Okada and Kazuaki Nakajima and Sho Tanosaki and Shota Someya and Akinori Hirano and Shinji Kawaguchi and Eiji Kobayashi and Keiichi Fukuda",
note = "Funding Information: The authors thank the Center for iPSC Research and Application, Kyoto University, for providing the 253G4, 201B7, and FfI14 hiPSC lines. This work was supported by the Highway Program for Realization of Regenerative Medicine from the Japan Science and Technology Agency (no. 16bm0504006h0006 to K.F.) and partly supported by the Support Program to Break the Bottlenecksat R&D for Accelerating the Practical Use of Health Research Outcomes (no. 34201006-01 to K.F.), a Japan Society for the Promotion of Science JSPS KAKENHI grant (no. 16K09507 to J.F., no. 15H04684 to J.K., and no. 15K09098 to H.K.), and the SENSHIN Medical Research Foundation (to S. Tohyama). The authors are grateful to Satoru Okamoto, Yusuke Natsume, and Kotoe Terasawa for providing the medium (Ajinomoto Co., Inc.) and to Yoshihide Miyatani and Akira Yamamoto for adjusting the incubators with the AGV system (Taitec Corporation). K.F., who is a co-founder of Heartseed, Inc., has equity in Heartseed, Inc. Funding Information: The authors thank the Center for iPSC Research and Application, Kyoto University, for providing the 253G4, 201B7, and FfI14 hiPSC lines. This work was supported by the Highway Program for Realization of Regenerative Medicine from the Japan Science and Technology Agency (no. 16bm0504006h0006 to K.F.) and partly supported by the Support Program to Break the Bottlenecks at R&D for Accelerating the Practical Use of Health Research Outcomes (no. 34201006-01 to K.F.), a Japan Society for the Promotion of Science JSPS KAKENHI grant (no. 16K09507 to J.F., no. 15H04684 to J.K., and no. 15K09098 to H.K.), and the SENSHIN Medical Research Foundation (to S. Tohyama). The authors are grateful to Satoru Okamoto, Yusuke Natsume, and Kotoe Terasawa for providing the medium (Ajinomoto Co., Inc.) and to Yoshihide Miyatani and Akira Yamamoto for adjusting the incubators with the AGV system (Taitec Corporation). K.F., who is a co-founder of Heartseed, Inc., has equity in Heartseed, Inc. Publisher Copyright: {\textcopyright} 2017 The Authors",
year = "2017",
month = nov,
day = "14",
doi = "10.1016/j.stemcr.2017.08.025",
language = "English",
volume = "9",
pages = "1406--1414",
journal = "Stem Cell Reports",
issn = "2213-6711",
publisher = "Cell Press",
number = "5",
}