Generation and Functional Characterization of Cadaver-Derived iPSC-Cardiomyocytes: A Feasibility Study Toward Functional Molecular Autopsy in Sudden Cardiac Death

Abstract

Sudden cardiac death (SCD) accounts for a substantial share of deaths in young individuals. Despite postmortem pathological, toxicological, and forensic examination and molecular autopsy, many cases remain unexplained, highlighting the need for complementary approaches to investigate the functional consequences of genetic variants identified after death. In this study, we describe a methodology for generating cardiomyocytes from human cadavers and demonstrate their functional viability. Achilles tendons were harvested from seven cadavers, all younger than 40 years of age, that had been preserved in a morgue and were subject to forensic autopsies. Fibroblasts were successfully cultured in six cases and reprogrammed into cadaver-derived induced pluripotent stem cells (CdiPSCs) in three cases. The CdiPSCs exhibited a stable pluripotent phenotype and were differentiated into cardiomyocytes (CdiPSC-CMs) positive for cardiac markers. Electrophysiological and optical characterization demonstrated functional cellular properties, including spontaneous and evoked action potentials, voltage-gated sodium currents, calcium wave propagation, and re-entrant rotor activity. CdiPSC-CMs also responded to β-adrenergic stimulation, demonstrating preservation of key excitable and signaling properties. Together, these findings establish the technical feasibility of recovering viable and functionally responsive cardiomyocytes from individuals post-death. This study provides a proof-of-concept platform for future functional studies of patient-specific genetic variants identified by molecular autopsy and, ultimately, for assessing their potential pathogenicity.