title: Multiparameter in vitro assessment of compound effects on cardiomyocyte physiology using iPSC cells.
abstract: A large percentage of drugs fail in clinical studies due to cardiac toxicity; thus, development of sensitive in vitro assays that can evaluate potential adverse effects on cardiomyocytes is extremely important for drug development. Human cardiomyocytes derived from stem cell sources offer more clinically relevant cell-based models than those presently available. Human-induced pluripotent stem cell-derived cardiomyocytes are especially attractive because they express ion channels and demonstrate spontaneous mechanical and electrical activity similar to adult cardiomyocytes. Here we demonstrate techniques for measuring the impact of pharmacologic compounds on the beating rate of cardiomyocytes with ImageXpress Micro and FLIPR Tetra systems. The assays employ calcium-sensitive dyes to monitor changes in Ca(2+) fluxes synchronous with cell beating, which allows monitoring of the beat rate, amplitude, and other parameters. We demonstrate here that the system is able to detect concentration-dependent atypical patterns caused by hERG inhibitors and other ion channel blockers. We also show that both positive and negative chronotropic effects on cardiac rate can be observed and IC(50) values determined. This methodology is well suited for safety testing and can be used to estimate efficacy and dosing of drug candidates prior to clinical studies.
authors.affiliation: Molecular Devices, LLC, Sunnyvale, CA, USA.
authors.forename: Oksana
authors.initials: O
authors.lastname: Sirenko
authors.forename: Carole
authors.initials: C
authors.lastname: Crittenden
authors.forename: Nick
authors.initials: N
authors.lastname: Callamaras
authors.forename: Jayne
authors.initials: J
authors.lastname: Hesley
authors.forename: Yen-Wen
authors.initials: YW
authors.lastname: Chen
authors.forename: Carlos
authors.initials: C
authors.lastname: Funes
authors.forename: Ivan
authors.initials: I
authors.lastname: Rusyn
authors.forename: Blake
authors.initials: B
authors.lastname: Anson
authors.forename: Evan F
authors.initials: EF
authors.lastname: Cromwell
mesh_terms: D020013:Calcium Signaling / Q000187:drug effects; D002316:Cardiotonic Agents / Q000494:pharmacology; D054715:Cardiotoxins / Q000494:pharmacology; D002478:Cells, Cultured; D003864:Depression, Chemical; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000502:physiology; D008856:Microscopy, Fluorescence; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013268:Stimulation, Chemical; D059008:Time-Lapse Imaging
chemical_list: D002316:Cardiotonic Agents; D054715:Cardiotoxins