Article abstract


Nature Methods 7, 61 - 66 (2010)
Published online: 29 November 2009 | doi:10.1038/nmeth.1403

Nongenetic method for purifying stem cell–derived cardiomyocytes

Fumiyuki Hattori1,2, Hao Chen1,3, Hiromi Yamashita1, Shugo Tohyama1,3, Yu-suke Satoh1,4, Shinsuke Yuasa1, Weizhen Li1, Hiroyuki Yamakawa1,3, Tomofumi Tanaka1,2, Takeshi Onitsuka1,3, Kenichiro Shimoji1,3, Yohei Ohno1,3, Toru Egashira1,3, Ruri Kaneda1, Mitsushige Murata1,3, Kyoko Hidaka5, Takayuki Morisaki5, Erika Sasaki6, Takeshi Suzuki4, Motoaki Sano1, Shinji Makino1, Shinzo Oikawa2 & Keiichi Fukuda1


Several applications of pluripotent stem cell (PSC)-derived cardiomyocytes require elimination of undifferentiated cells. A major limitation for cardiomyocyte purification is the lack of easy and specific cell marking techniques. We found that a fluorescent dye that labels mitochondria, tetramethylrhodamine methyl ester perchlorate, could be used to selectively mark embryonic and neonatal rat cardiomyocytes, as well as mouse, marmoset and human PSC-derived cardiomyocytes, and that the cells could subsequently be enriched (>99% purity) by fluorescence-activated cell sorting. Purified cardiomyocytes transplanted into testes did not induce teratoma formation. Moreover, aggregate formation of PSC-derived cardiomyocytes through homophilic cell-cell adhesion improved their survival in the immunodeficient mouse heart. Our approaches will aid in the future success of using PSC-derived cardiomyocytes for basic and clinical applications.

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  1. Department of Regenerative Medicine and Advanced Cardiac Therapeutics, Keio University School of Medicine, Tokyo, Japan.
  2. Asubio Pharma Co., Ltd., Osaka, Japan.
  3. Division of Cardiology, Department of Medicine, Keio University School of Medicine, Tokyo, Japan.
  4. Division of Basic Biological Sciences, Faculty of Pharmacy, Keio University, Tokyo, Japan.
  5. Department of Bioscience, National Cardiovascular Center Research Institute, Osaka, Japan.
  6. Laboratory of Applied Developmental Biology, Marmoset Research Department, Central Institute for Experimental Animals, Kanagawa, Japan.

Correspondence to: Keiichi Fukuda1 e-mail: kfukuda@sc.itc.keio.ac.jp



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