Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings
Clark, Alexander P; Clerx, Michael; Wei, Siyu; Lei, Chon Lok; de Boer, Teun P; Mirams, Gary R; Christini, David J; Krogh-Madsen, Trine
(2023) Europace : European pacing, arrhythmias, and cardiac electrophysiology : journal of the working groups on cardiac pacing, arrhythmias, and cardiac cellular electrophysiology of the European Society of Cardiology, volume 25, issue 9
(Article)
Abstract
Aims: Human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) have become an essential tool to study arrhythmia mechanisms. Much of the foundational work on these cells, as well as the computational models built from the resultant data, has overlooked the contribution of seal-leak current on the immature and heterogeneous phenotype that has
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come to define these cells. The aim of this study is to understand the effect of seal-leak current on recordings of action potential (AP) morphology. Methods and results: Action potentials were recorded in human iPSC-CMs using patch clamp and simulated using previously published mathematical models.
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Keywords: Arrhythmias, Computer simulation, Induced pluripotent stem cells, Ion channels, Patch clamp, Cardiology and Cardiovascular Medicine, Physiology (medical), Journal Article
ISSN: 1099-5129
Publisher: Oxford University Press
Note: Publisher Copyright: ©. We show that compensation of this leak current is difficult due to challenges with obtaining accurate measures of R during an experiment. Using simulation, we show that R measures (i) change during an experiment, invalidating the use of pre-rupture values, and (ii) are polluted by the presence of transmembrane currents at every voltage. Finally, we posit that the background sodium current in baseline iPSC-CM models imitates the effects of seal-leak current and is increased to a level that masks the effects of seal-leak current on iPSC-CMs. Conclusion: Based on these findings, we make recommendations to improve iPSC-CM AP data acquisition, interpretation, and model-building. Taking these recommendations into account will improve our understanding of iPSC-CM physiology and the descriptive ability of models built from such data.
(Peer reviewed)
