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Author Notes:

To whom correspondence should be addressed: Chunhui Xu; Email: chunhui.xu@emory.edu

Subjects:

Research Funding:

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, and grants from the National Institutes of Health (R21HL118454), and R21HL123928); and a grant from CASIS (GA-2014-126).

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Cell Biology
  • Cardiomyocytes
  • Differentiation
  • Flow cytometry analysis
  • Growth factors
  • Immunocytochemical analysis
  • Pluripotent stem cells
  • qRT-PCR
  • Serum-free medium
  • MODULATION
  • MATURATION
  • ENRICHMENT
  • MYOCYTES
  • DEVELOP

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

Tools:

Journal Title:

Methods in Molecular Biology

Volume:

Volume 1299

Publisher:

, Pages 115-131

Type of Work:

Article | Post-print: After Peer Review

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.

Copyright information:

© Springer Science+Business Media New York 2015.

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