title: Methods for the derivation and use of cardiomyocytes from human pluripotent stem cells.
abstract: The availability of human cardiomyocytes derived from embryonic stem cells (ESCs) has generated -considerable excitement, as these cells are an excellent model system for studying myocardial development and may have eventual application in cell-based cardiac repair. Cardiomyocytes derived from the related induced pluripotent stem cells (iPSCs) have similar properties, but also offer the prospects of patient-specific disease modeling and cell therapies. Unfortunately, the methods by which cardiomyocytes have been historically generated from pluripotent stem cells are unreliable and typically result in preparations of low cardiac purity (typically <1% cardiomyocytes). We detail here the methods for a recently reported directed cardiac differentiation protocol, which involves the serial application of two growth factors known to be involved in early embryonic heart development, activin A, and bone morphogenetic protein-4 (BMP-4). This protocol reliably yields preparations of 30-60% cardiomyocytes, which can then be further enriched to >90% cardiomyocytes using straightforward physical methods.
authors.affiliation: Laflamme Lab, Pathology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
authors.forename: Wei-Zhong
authors.initials: WZ
authors.lastname: Zhu
authors.forename: Benjamin
authors.initials: B
authors.lastname: Van Biber
authors.forename: Michael A
authors.initials: MA
authors.lastname: Laflamme
mesh_terms: D028341:Activins / Q000494:pharmacology; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003094:Collagen / Q000494:pharmacology; D015925:Cryopreservation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004338:Drug Combinations; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011205:Povidone; D011509:Proteoglycans / Q000494:pharmacology; D012822:Silicon Dioxide
chemical_list: D055415:Bone Morphogenetic Protein 4; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C422405:activin A; D028341:Activins; C059485:matrigel; C016039:Percoll; D012822:Silicon Dioxide; D003094:Collagen; D011205:Povidone