Chapter title |
Efficient Differentiation of Cardiomyocytes from Human Pluripotent Stem Cells with Growth Factors
|
---|---|
Chapter number | 9 |
Book title |
Cardiomyocytes
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2572-8_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2571-1, 978-1-4939-2572-8
|
Authors |
Jha, Rajneesh, Xu, Ren-He, Xu, Chunhui, Rajneesh Jha, Ren-He Xu, Chunhui Xu |
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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