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Nat Methods. 2014 Aug;11(8):855-60. doi: 10.1038/nmeth.2999. Epub 2014 Jun 15.

Chemically defined generation of human cardiomyocytes.

Author information

1
1] Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California, USA. [2] Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA. [3] Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California, USA.
2
Department of Applied Physics, Stanford University School of Medicine, Stanford, California, USA.
3
Department of Chemistry, Stanford University School of Medicine, Stanford, California, USA.
4
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California, USA.

Abstract

Existing methods for human induced pluripotent stem cell (hiPSC) cardiac differentiation are efficient but require complex, undefined medium constituents that hinder further elucidation of the molecular mechanisms of cardiomyogenesis. Using hiPSCs derived under chemically defined conditions on synthetic matrices, we systematically developed an optimized cardiac differentiation strategy, using a chemically defined medium consisting of just three components: the basal medium RPMI 1640, L-ascorbic acid 2-phosphate and rice-derived recombinant human albumin. Along with small molecule-based induction of differentiation, this protocol produced contractile sheets of up to 95% TNNT2(+) cardiomyocytes at a yield of up to 100 cardiomyocytes for every input pluripotent cell and was effective in 11 hiPSC lines tested. This chemically defined platform for cardiac specification of hiPSCs will allow the elucidation of cardiomyocyte macromolecular and metabolic requirements and will provide a minimal system for the study of maturation and subtype specification.

PMID:
24930130
PMCID:
PMC4169698
DOI:
10.1038/nmeth.2999
[Indexed for MEDLINE]
Free PMC Article
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