Abstract Wed145: Multiplexed Electrophysiological, Contractile, and Morphological Analysis for High-Throughput In Vitro Cardiotoxicity Testing and Drug Development

Despite advances in drug development, the lack of high-throughput cardiac technologies limits the efficient assessment of in vitro functional and structural cardiotoxicity. This gap hinders the rapid and comprehensive evaluation of drug-induced cardiac effects, posing challenges for early detection and risk mitigation. Moreover, most systems solely assay electrophysiological activity, largely overlooking structural and contractile changes in cardiomyocytes and supporting cells that contribute to disease and toxicity. CytoTronics’ Pixel electrical imaging platform addresses this critical need by enabling high-throughput (up to eight 96- or 384-well plates), non-invasive, and label-free live-cell multiparametric analysis. Featuring a high-density electrode array with 400 µm spatial resolution, the platform simultaneously captures electrophysiological and contractility data for functional assessments of electrogenic cells. Additionally, the same electrodes at 12.5 µm resolution measure over 20 functional and morphological parameters—including tissue barrier integrity, cell-surface attachment, and motility—through advanced field-based impedance analysis. With real-time measurements spanning minutes to hours, the platform generates electrical images, time-lapse videos, time-course line plots, and high dimensional principal component analyses, offering deep longitudinal insights into cellular dynamics. In this study, we tracked multiplexed measurements of human iPSC-derived cardiomyocytes grown for seven days following treatment with cardiotoxic and positive inotropic compounds. The analysis revealed distinct effects, with functionally cardiotoxic compounds significantly altering field potential duration, structurally cardiotoxic compounds impacting impedance measurements, and positive inotropic compounds affecting contractility. Cells were assessed for up to one week post-treatment to capture longitudinal, chronic responses to compound treatment. These findings highlight the Pixel platform’s ability to provide comprehensive, high-throughput functional and structural cardiotoxicity assessments, offering a valuable tool for drug safety evaluation and cardiovascular research.