Abstract 16578: Scalable, Reproducible, and Economical Method for Producing Cardiomyocytes From Human Induced Pluripotent Stem Cells
Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are considered to be a promising tool for various purposes, e.g. regenerative medicine, drug discovery, and safety pharmacology. Many researchers have reported various protocols for inducing cardiac differentiation from hiPSC. For above-mentioned purposes, ideal protocol for cardiac differentiation should be scalable (3D-culture preferred), inexpensive (use of small molecule preferred), and highly reproducible (defined factor preferred).
Here, we report an innovative protocol of cardiac differentiation, which is based on widely used protocol from Gordon Keller’s lab that uses various cytokines in a 3D-culture system (Yang L et al. Nature 2008). In the new protocol, costs for inducing cardiac differentiation was reduced by 96% with improved differentiation efficiency and high interclonal reproducibility.
According to our previous report (Naito AT et al. PNAS 2006), we tested whether appropriate activation and inhibition of Wnt signaling can replace the cytokines used for cardiac differentiation. We found that activator of Wnt signaling can by itself replace the cytokine that was used for mesoderm induction (Activin A and BMP). Although inhibitor of Wnt signaling alone could not replace the cytokines that was used after mesoderm induction (Dkk-1, bFGF, and VEGF), robust cardiac differentiation was observed when Wnt inhibitor was used in combination with inhibitor of Tgfb signaling and a steroid hormone. We have also developed a differentiation medium that is consisted of defined and inexpensive factors. The total cost for inducing cardiac differentiation was cut from 1,376,250 yen (~$13,762) to 57,150 yen (~$571) in a 1L scale. Our new protocol and culture medium would dramatically reduce the cost for producing cardiomyocytes from iPSCs and would promote utilization of iPSC-derived cardiomyocytes for regenerative medicine, drug discovery and safety pharmacology.
Author Disclosures: M. Ito: None. R. Takizawa: None. N. Igarashi: None. M. Naito: None. I. Komuro: Research Grant; Modest; SHIONOGI & CO., LTD., Glaxo Smith Kline., Sanofi K.K., Genzyme Japan K.K., Mitsubishi Tanabe Pharma Corporation, Eli Lilly Japan K.K., Pfizer Japan Inc., Bristol-Myers Squibb Company. Other Research Support; Modest; Takeda Pharmaceutical Company Limited., Nippon Boehringer Ingelheim Co., Ltd., MSD K.K., SHIONOGI & CO., LTD., Astellas Pharma Inc., Mitsubishi Tanabe Pharma Corporation, Novartis Pharma K.K., Kyowa Hakko Kirin Co.,Ltd., TEIJIN PHARMA LIMITED., DAIICHI SANKYO COMPANY, LIMITED, Otsuka Pharmaceutical Co., Ltd.. Honoraria; Modest; Takeda Pharmaceutical Company Limited., Nippon Boehringer Ingelheim Co., Ltd., SHIONOGI & CO., LTD., Astellas Pharma Inc., Novartis Pharma K.K., DAIICHI SANKYO COMPANY, LIMITED. Consultant/Advisory Board; Modest; Dainippon Sumitomo Pharma Co., Ltd., ONO PHARMACEUTICAL CO., LTD. A. Naito: Research Grant; Modest; SHIONOGI & CO., LTD., Genzyme Japan K.K., Mitsubishi Tanabe Pharma Corporation, ONO PHARMACEUTICAL CO., LTD., DAIICHI SANKYO COMPANY, LIMITED. Other Research Support; Modest; ONO PHARMACEUTICAL CO., LTD., DAIICHI SANKYO COMPANY, LIMITED. Honoraria; Modest; Nippon Boehringer Ingelheim Co., Ltd., Novartis Pharma K.K., KOWA PHARMACEUTICAL CO., LTD..
- © 2014 by American Heart Association, Inc.