title: Derivation of highly purified cardiomyocytes from human induced pluripotent stem cells using small molecule-modulated differentiation and subsequent glucose starvation.
abstract: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have become an important cell source to address the lack of primary cardiomyocytes available for basic research and translational applications. To differentiate hiPSCs into cardiomyocytes, various protocols including embryoid body (EB)-based differentiation and growth factor induction have been developed. However, these protocols are inefficient and highly variable in their ability to generate purified cardiomyocytes. Recently, a small molecule-based protocol utilizing modulation of Wnt/β-Catenin signaling was shown to promote cardiac differentiation with high efficiency. With this protocol, greater than 50%-60% of differentiated cells were cardiac troponin-positive cardiomyocytes were consistently observed. To further increase cardiomyocyte purity, the differentiated cells were subjected to glucose starvation to specifically eliminate non-cardiomyocytes based on the metabolic differences between cardiomyocytes and non-cardiomyocytes. Using this selection strategy, we consistently obtained a greater than 30% increase in the ratio of cardiomyocytes to non-cardiomyocytes in a population of differentiated cells. These highly purified cardiomyocytes should enhance the reliability of results from human iPSC-based in vitro disease modeling studies and drug screening assays.
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine.
authors: Arun
authors: A
authors: Sharma
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine.
authors: Guang
authors: G
authors: Li
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine.
authors: Kuppusamy
authors: K
authors: Rajarajan
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine.
authors: Ryoko
authors: R
authors: Hamaguchi
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine.
authors: Paul W
authors: PW
authors: Burridge
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine; Institute of Stem Cell Biology and Regenerative Medicine, Cardiovascular Medicine Division, Department of Medicine, Child Health Research Institute, Stanford University School of Medicine; smwu@stanford.edu.
authors: Sean M
authors: SM
authors: Wu
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D003470:Culture Media; D005947:Glucose / Q000172:deficiency* / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D003470:Culture Media; D005947:Glucose