Chemically Defined Culture and Cardiomyocyte Differentiation of Human Pluripotent Stem Cells

Paul W. Burridge, Alexandra Holmström, Joseph C. Wu

Research output: Contribution to journalArticlepeer-review

131 Scopus citations

Abstract

Since the first discovery that human pluripotent stem cells (hPS cells) can differentiate to cardiomyocytes, efforts have been made to optimize the conditions under which this process occurs. One of the most effective methodologies to optimize this process is reductionist simplification of the medium formula, which eliminates complex animal-derived components to help reveal the precise underlying mechanisms. Here we describe our latest, cost-effective and efficient methodology for the culture of hPS cells in the pluripotent state using a modified variant of chemically defined E8 medium. We provide exact guidelines for cell handling under these conditions, including non-enzymatic EDTA passaging, which have been optimized for subsequent cardiomyocyte differentiation. We describe in depth the latest version of our monolayer chemically defined small molecule differentiation protocol, including metabolic selection–based cardiomyocyte purification and the addition of triiodothyronine to enhance cardiomyocyte maturation. Finally, we describe a method for the dissociation of hPS cell–derived cardiomyocytes, cryopreservation, and thawing.

Original languageEnglish (US)
Pages (from-to)21.3.1-21.3.15
JournalCurrent protocols in human genetics
Volume87
Issue number1
DOIs
StatePublished - Oct 1 2015

Funding

This work was funded by NIH K99 HL121177 and American Heart Association Grant 14BGIA 20480329 (to P.W.B.) and Postdoctoral Fellowship from Vetenskapsrådet (to A.H.) and American Heart Association Established Investigator Award, NIH grant R01 HL113006, NIH R01 HL123968, NIH R01 HL125627, and NIH R24 HL117756 (to J.C.W).

Keywords

  • cardiac
  • cardiomyocyte
  • chemically defined
  • differentiation
  • human induced pluripotent stem cells
  • monolayer

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

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