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Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings
Alexander P Clark1; Michael Clerx2; Siyu Wei3; Chon Lok Lei4,5; Teun P de Boer6; Gary R Mirams2; David J Christini1,3; Trine Krogh-Madsen7,8
2023-08-08
Source PublicationEUROPACE
ISSN1099-5129
Volume25Issue:9Pages:euad243
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.

KeywordArrhythmias Computer Simulation Induced Pluripotent Stem Cells Ion Channels Patch Clamp
DOI10.1093/europace/euad243
URLView the original
Indexed BySCIE
Language英語English
WOS Research AreaCardiovascular System & Cardiology
WOS SubjectCardiac & Cardiovascular Systems
WOS IDWOS:001054221500006
PublisherNLM (Medline)
Scopus ID2-s2.0-85168803290
Fulltext Access
Citation statistics
Document TypeJournal article
CollectionFaculty of Health Sciences
Institute of Translational Medicine
DEPARTMENT OF BIOMEDICAL SCIENCES
Corresponding AuthorTrine Krogh-Madsen
Affiliation1.Department of Biomedical Engineering, Cornell University, Ithaca, United States
2.Centre for Mathematical Medicine and Biology, School of Mathematical Sciences, University of Nottingham, Nottingham, United Kingdom
3.Department of Physiology and Pharmacology, SUNY Downstate Health Sciences University, Brooklyn, United States
4.Institute of Translational Medicine, Faculty of Health Sciences, University of Macau, Macau, China
5.Department of Biomedical Sciences, Faculty of Health Sciences, University of Macau, Macau, China
6.Department of Medical Physiology, Division of Heart and Lungs, University Medical Center Utrecht, Netherlands
7.Department of Physiology and Biophysics, Weill Cornell Medicine, 1300 York Avenue, Box 75, Room C501D, New York, 10065 NY, USA
8.Institute for Computational Biomedicine, Weill Cornell Medicine, 1300 York Avenue, Box 75, Room C501D, New York, 10065 NY, USA
Recommended Citation
GB/T 7714
Alexander P Clark,Michael Clerx,Siyu Wei,et al. Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings[J]. EUROPACE, 2023, 25(9), euad243.
APA Alexander P Clark., Michael Clerx., Siyu Wei., Chon Lok Lei., Teun P de Boer., Gary R Mirams., David J Christini., & Trine Krogh-Madsen (2023). Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings. EUROPACE, 25(9), euad243.
MLA Alexander P Clark,et al."Leak current, even with gigaohm seals, can cause misinterpretation of stem cell-derived cardiomyocyte action potential recordings".EUROPACE 25.9(2023):euad243.
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