title: High-throughput drug profiling with voltage- and calcium-sensitive fluorescent probes in human iPSC-derived cardiomyocytes.
abstract: Cardiomyocytes derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (hiPSCs) are increasingly used for in vitro assays and represent an interesting opportunity to increase the data throughput for drug development. In this work, we describe a 96-well recording of synchronous electrical activities from spontaneously beating hiPSC-derived cardiomyocyte monolayers. The signal was obtained with a fast-imaging plate reader using a submillisecond-responding membrane potential recording assay, FluoVolt, based on a newly derived voltage-sensitive fluorescent dye. In our conditions, the toxicity of the dye was moderate and compatible with episodic recordings for >3 h. We show that the waveforms recorded from a whole well or from a single cell-sized zone are equivalent and make available critical functional parameters that are usually accessible only with gold standard techniques like intracellular microelectrode recording. This approach allows accurate identification of the electrophysiological effects of reference drugs on the different phases of the cardiac action potential as follows: fast depolarization (lidocaine), early repolarization (nifedipine, Bay K8644, and veratridine), late repolarization (dofetilide), and diastolic slow depolarization (ivabradine). Furthermore, the data generated with the FluoVolt dye can be pertinently complemented with a calcium-sensitive dye for deeper characterization of the pharmacological responses. In a semiautomated plate reader, the two probes used simultaneously in 96-well plates provide an easy and powerful multiparametric assay to rapidly and precisely evaluate the cardiotropic profile of compounds for drug discovery or cardiac safety.
authors.affiliation: Laboratoire SERVIER de Chemogénétique, Institut du Cerveau et de la Moelle, Hôpital de la Salpétrière, Paris, France; Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France stephane.bedut.vwr@servier.com.
authors.forename: Stephane
authors.initials: S
authors.lastname: Bedut
authors.affiliation: Laboratoire SERVIER de Chemogénétique, Institut du Cerveau et de la Moelle, Hôpital de la Salpétrière, Paris, France;
authors.forename: Christine
authors.initials: C
authors.lastname: Seminatore-Nole
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Veronique
authors.initials: V
authors.lastname: Lamamy
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Sarah
authors.initials: S
authors.lastname: Caignard
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Jean A
authors.initials: JA
authors.lastname: Boutin
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Nosjean
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Stephan
authors.affiliation: Pôle d'Expertise Biotechnologie, Chimie & Biologie, Institut de Recherches SERVIER, Croissy-sur-Seine, France.
authors.forename: Francis
authors.initials: F
authors.lastname: Coge
mesh_terms: D000200:Action Potentials / Q000187:drug effects*; D057205:Automation, Laboratory; D020013:Calcium Signaling / Q000187:drug effects*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005456:Fluorescent Dyes / Q000378:metabolism* / Q000633:toxicity; D057166:High-Throughput Screening Assays; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009200:Myocardial Contraction / Q000187:drug effects*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D012815:Signal Processing, Computer-Assisted; D013997:Time Factors
chemical_list: D005456:Fluorescent Dyes
keywords: 96-well plate; cardiomyocytes; cell signaling; induced pluripotent stem cells; membrane potentials