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

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 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. Our in silico and in vitro studies demonstrate how seal–leak current depolarizes APs, substantially affecting their morphology, even with seal resistances (Rseal) above 1 GΩ. We show that compensation of this leak current is difficult due to challenges with obtaining accurate measures of Rseal during an experiment. Using simulation, we show that Rseal 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.

Citation

Clark, A. P., Clerx, M., Wei, S., Lei, C. L., de Boer, T. P., Mirams, G. R., …Krogh-Madsen, T. (2023). Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings. EP-Europace, 25(9), Article euad243. https://doi.org/10.1093/europace/euad243

• Journal Article Type: Article
• Acceptance Date: Jun 18, 2023
• Online Publication Date: Aug 8, 2023
• Publication Date: 2023-09
• Deposit Date: Aug 14, 2023
• Publicly Available Date: Aug 14, 2023
• Journal: EP-Europace
• Print ISSN: 1099-5129
• Electronic ISSN: 1532-2092
• Publisher: Oxford University Press
• Peer Reviewed: Peer Reviewed
• Volume: 25
• Issue: 9
• Article Number: euad243
• DOI: https://doi.org/10.1093/europace/euad243
• Keywords: Induced pluripotent stem cells, Patch clamp, Arrhythmias, Ion channels, Computer simulation
• Public URL: https://nottingham-repository.worktribe.com/output/24148151
• Publisher URL: https://academic.oup.com/europace/article/25/9/euad243/7239258?