hiPSC-CM Electrophysiology: Impact of Temporal Changes and Study Parameters on Experimental Reproducibility

Authors

Devon Guerrelli, Children's National Heart Institute, Children's National Hospital, Washington, DC; Sheikh Zayed Institute for Pediatric Surgical Innovation, Children's National Research Institute, Washington, DC, Washington, D.C., United States.
Jenna Pressman, Children's National Research Institute, Washington, United States.
Shatha Salameh, Children's National, United States.
Nikki G. Posnack, Sheikh Zayed Institute for Pediatric Surgical Innovation, Children's National Hospital; Department of Pediatrics, George Washington University School of Medicine, Washington, D.C., DC, United States.

Document Type

Journal Article

Publication Date

5-10-2024

Journal

American journal of physiology. Heart and circulatory physiology

DOI

10.1152/ajpheart.00631.2023

Keywords

CiPA; Electrophysiology; hiPSC-CM; microelectrode array; reproducibility

Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are frequently used for preclinical cardiotoxicity testing and remain an important tool for confirming model-based predictions of drug effects in accordance with Comprehensive Proarrhythmia Assay (CiPA). Despite the considerable benefits hiPSC-CMs provide, concerns surrounding experimental reproducibility have emerged. We investigated the effects of temporal changes and experimental parameters on hiPSC-CM electrophysiology. iCell cardiomyocytes were cultured and biosignals were acquired using a microelectrode array (MEA) system (1-14 days). Continuous recordings revealed a 22.6% increase in the beating rate and 7.7% decrease in the field potential duration (FPD) during a 20-minute equilibration period. Location specific differences across a multiwell plate were also observed, with iCell cardiomyocytes in the outer rows beating 8.8 beats per minute (BPM) faster than the inner rows. Cardiac endpoints were also impacted by cell culture duration; from 2-14 days the beating rate decreased (-12.7 BPM), FPD lengthened (+257 ms), and spike amplitude increased (+3.3 mV). Cell culture duration (4-10 days) also impacted cardiomyocyte drug responsiveness (E-4031, nifedipine, isoproterenol). qRT-PCR results suggest that daily variations in cardiac metrics may be linked to the continued maturation of hiPSC-CMs in culture (2-30 days). Daily experiments were also repeated using a second cell line (Cor.4U). Collectively, our study highlights multiple sources of variability to consider and address when performing hiPSC-CM MEA studies. To improve reproducibility and data interpretation, MEA-based studies should establish a standardized protocol and report key experimental conditions (e.g., cell line, culture time, equilibration time, electrical stimulation settings, raw data values).

APA Citation

Guerrelli, Devon; Pressman, Jenna; Salameh, Shatha; and Posnack, Nikki G., "hiPSC-CM Electrophysiology: Impact of Temporal Changes and Study Parameters on Experimental Reproducibility" (2024). GW Authored Works. Paper 4955.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/4955

Department

Pediatrics