Chronic Electrophysiology Captures Progressive Variability in DNA-Damaged Cardiac Networks

Abstract Electrophysiological recordings provide a key functional readout of cardiac health. Most in vitro studies rely on acute measurements rather than recording evolving functional changes. Here, we implement a microelectrode array (MEA) based system for chronic electrophysiological monitoring of human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) monolayers. Electrophysiology recordings were maintained for up to 40 days, enabling longitudinal assessment of field potential amplitude, field potential duration, beat frequency, conduction velocity, and waveform morphology. Untreated iPSC-CM monolayers exhibited stable electrophysiological behavior throughout chronic culture. Following exposure to the cardiotoxic chemotherapeutic doxorubicin, chronic recordings captured progressive deterioration in electrical activity, including reduced field potential amplitude, altered repolarization dynamics, increased beat-to-beat variability, and reduced conduction that persisted after drug removal. Electrode-resolved analyses further revealed pronounced progressive spatial variability, highlighting localized responses that would be obscured by population-averaged measurements. Endpoint assays indicated reduced calcium transient amplitude and altered calcium reuptake, decreased mitochondrial membrane potential, and loss of connexin 43, findings consistent with the impaired electrical activity observed in chronic recordings. Together, these results establish chronic electrophysiological monitoring as a powerful approach for resolving the spatiotemporal progression of cardiotoxicity and other dynamic cardiac processes in vitro .