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Efficient Differentiation of Cardiomyocytes from Human Pluripotent Stem Cells with Growth Factors

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Cardiomyocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1299))

Abstract

Human pluripotent stem cells have tremendous replicative capacity and demonstrated potential to generate functional cardiomyocytes. These cardiomyocytes represent a promising source for cell replacement therapy to treat heart disease and may serve as a useful tool for drug discovery and disease modeling. Efficient cardiomyocyte differentiation, a prerequisite for the application of stem cell-derived cardiomyocytes, can be achieved with a growth factor-guided method. Undifferentiated cells are sequentially treated with activin A and BMP4 in a serum-free and insulin-free medium and then maintained in a serum-free medium with insulin. This method yields as much as >75 % cardiomyocytes in the differentiation culture within 2 weeks, and the beating cardiomyocytes have expected molecular, cellular, and electrophysiological characteristics. In this chapter, we describe in detail the differentiation protocol and follow-up characterization focusing on immunocytochemistry, quantitative RT-PCR, and flow cytometry analysis.

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Acknowledgments

The C. Xu laboratory gratefully acknowledges the funding from the Children’s Pediatric Research Trust from Emory Children’s Pediatric Research Center; the funding from the National Heart, Lung, and Blood Institute; National Institutes of Health, under Contract No. HHSN268201000043C; grants from the National Institutes of Health (R21HL118454 and R21HL123928); and a grant from CASIS (GA-2014-126).

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Authors and Affiliations

  1. Department of Pediatrics, Emory University School of Medicine and Children’s Healthcare of Atlanta, 2015 Uppergate Road, Atlanta, GA, 30322, USA

    Rajneesh Jha & Chunhui Xu

  2. Faculty of Health Sciences, University of Macau, Taipa, Macau, China

    Ren-He Xu

  3. The Parker H. Petit Institute for Bioengineering and Bioscience, Atlanta, GA, 30332, USA

    Chunhui Xu

Authors
  1. Rajneesh Jha
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  2. Ren-He Xu
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  3. Chunhui Xu
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Corresponding author

Correspondence to Chunhui Xu .

Editor information

Editors and Affiliations

  1. Rochester Institute of Technology, Rochester, New York, USA

    Gary R. Skuse

  2. Rochester Institute of Technology, Rochester, New York, USA

    Maureen C. Ferran

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Jha, R., Xu, RH., Xu, C. (2015). Efficient Differentiation of Cardiomyocytes from Human Pluripotent Stem Cells with Growth Factors. In: Skuse, G., Ferran, M. (eds) Cardiomyocytes. Methods in Molecular Biology, vol 1299. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2572-8_9

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  • DOI: https://doi.org/10.1007/978-1-4939-2572-8_9

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2571-1

  • Online ISBN: 978-1-4939-2572-8

  • eBook Packages: Springer Protocols

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