Automated Functional Cellular Analyses of Human iPS-derived Cardiomyocytes
Oksana Sirenko1, Nick Callamaras1, Jayne Hesley1, Xin Jiang1, Carole Crittenden1, Roger Tang1, Blake Anson2, and Evan F. Cromwell1
1Molecular Devices, Inc., 1311 Orleans Drive, Sunnyvale, CA 94089
2Cellular Dynamics International, 525 Science Drive, Stuite200, Madison, WI 53711
Human cardiomyocytes derived from stem cell sources can greatly accelerate the discovery of cardiac drugs and improve drug safety by offering more clinically relevant cell-based models than presently available. iPS-derived cardiomyocytes are especially attractive because they express ion channels and demonstrate beating and action potentials similar to primary cardiac cells. One emerging application for iPS-derived cardiomyocytes is for use as a model cell-based system for testing functional effects of ion channel blockers, GPCR antagonists, or other prospective drugs on cardiac contractility. This aspect, coupled with availability of such cells in large quantities, makes them useful for screening of lead compounds and important for reducing animal experimentation and cost of pre-clinical development.
Here we demonstrate live-cell assays for measuring the impact of pharmacological compounds on the rate and magnitude of beating cardiomyocytes using high content imaging. Spontaneously beating iPS-derived cardiomyocytes were cultivated in monolayer on 96w or 384w plates resulting in synchronized contractions. We developed a protocol that enables image acquisition and automatic determination of beating rate and magnitude of iPS-derived cardiomyocytes from time-lapse images of live cardiomyocytes. We demonstrate applications of these assays by measuring impact of norepinephrine, epinephrine, caffeine or acetylcholine on the beating rate. Tested reagents modulated the frequency of beating in line with their mode-of-action showing the functional expression of ß-adrenergic and acetylcholine receptors. We have also monitored other functional aspects underlying cardiomyocyte contractions including intracellular Ca2+ transient fluxes by using Ca2+ -sensitive fluorophores and presence of functional K+ and Ca2+ channels with automated patch clamp instruments.
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