title: Rapid cellular phenotyping of human pluripotent stem cell-derived cardiomyocytes using a genetically encoded fluorescent voltage sensor.
abstract: In addition to their promise in regenerative medicine, pluripotent stem cells have proved to be faithful models of many human diseases. In particular, patient-specific stem cell-derived cardiomyocytes recapitulate key features of several life-threatening cardiac arrhythmia syndromes. For both modeling and regenerative approaches, phenotyping of stem cell-derived tissues is critical. Cellular phenotyping has largely relied upon expression of lineage markers rather than physiologic attributes. This is especially true for cardiomyocytes, in part because electrophysiological recordings are labor intensive. Likewise, most optical voltage indicators suffer from phototoxicity, which damages cells and degrades signal quality. Here we present the use of a genetically encoded fluorescent voltage indicator, ArcLight, which we demonstrate can faithfully report transmembrane potentials in human stem cell-derived cardiomyocytes. We demonstrate the application of this fluorescent sensor in high-throughput, serial phenotyping of differentiating cardiomyocyte populations and in screening for drug-induced cardiotoxicity.
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Jordan S
authors: JS
authors: Leyton-Mange
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Robert W
authors: RW
authors: Mills
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Vincenzo S
authors: VS
authors: Macri
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Min Young
authors: MY
authors: Jang
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Faraz N
authors: FN
authors: Butte
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: Patrick T
authors: PT
authors: Ellinor
authors: Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors: David J
authors: DJ
authors: Milan
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: C576393:Arclight fusion protein; D015415:Biomarkers; D017077:Culture Media, Conditioned; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins