Abstract
Kinetic Image Cytometry™ (KIC™), in which living cells are cultured in multi-well dishes and imaged for electrophysiological transients (such as intracellular calcium or membrane potential) using automated digital microscopy/image analysis, represents an advance in the field of high content analysis. KIC methods, coupled with the use of human stem cell-derived cardiomyocytes (hSC-CMs), are potentially very useful for identifying proarrhythmia activity of test compounds, relevant to cardiosafety. In the present study, a panel of 40 compounds was tested with KIC to quantify the effects of the chemicals on the calcium transients associated with contraction in hSC-CMs, and the results compared to the effects of these compounds on the QT-interval from human electrocardiograms. The kinetics of the calcium transients quantified with KIC identified the chemicals that prolong (commonly associated with proarrhythmic activity) or shorten the QT interval with 100 % specificity and 94 % sensitivity. These data, along with data obtained in previous studies utilizing KIC and hSC-CMs, demonstrate that this assay platform represents the current “state-of-the-art” in vitro method for testing drug candidates for proarrhythmic tendencies.
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Acknowledgements
We would like to gratefully acknowledge Axiogenesis for contribution of the hES-CMs used in this study. Development of KIC instrumentation and methods and CyteSeer™ has been supported by several grants including the following: NIH/NHLBI FastTrack STTR R42HL086076 “Live cell and HCS assays to quantify production of cardiomyocytes from stem cells”; CIRM RT1-01143 “Optimization in the Identification, Selection and Induction of Maturation of Subtypes of Cardiomyocytes derived from Human Embryonic Stem Cells”; and NIH/NHLBI STTR R42HL112521 “Optogenetic Multiparametric Assay for HT Cardiotoxicity Testing.”
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Pfeiffer, E.R., Whittaker, R., Vega, R., Cerignoli, F., McDonough, P.M., Price, J.H. (2017). Kinetic Image Cytometry for Predicting Arrhythmias Using Human Stem Cell-Derived Cardiomyocytes. In: Clements, M., Roquemore, L. (eds) Stem Cell-Derived Models in Toxicology. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6661-5_8
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DOI: https://doi.org/10.1007/978-1-4939-6661-5_8
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