Model-Informed Characterization of Maturation-Related Electrophysiological States in Bioengineered hiPSC-CM Tissues

Teresa Diaz Jorda1, Manuel Antonio Oria Muriel2, Beatriz Trenor3, Marcos Latorre4, Jorge Sanchez5
1Ci2B, Universitat Politècnica de València, València, Spain, 2Ci2B, Universitat Politècnica de València, 3Universitat Politècnica de València, 4Ci2B, Universitat Politecnica de Valencia, 5Center for Research and Innovation in Bioengineering (Ci2B), Universitat Politecnica de Valencia


Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are widely used in cardiac tissue engineering, but their immature and heterogeneous electrophysiological phenotype remains a major limitation for functional tissue design and translational applications. Model-informed analysis may help bridge this gap by linking tissue-level measurements to the underlying cellular mechanisms that govern maturation-related behavior.

In this work, we combined optical mapping (OM) recordings and computational modeling to characterize the electrophysiological state of bioengineered hiPSC-CM tissues under two culture conditions: standard medium (SM) and hormonal treatment (HT). Voltage and calcium signals were extracted from OM recordings acquired after two weeks of culture. The Kernik hiPSC-CM model implemented in openCARP was calibrated to the experimental data using a genetic algorithm that optimized key ionic conductances to reproduce action potential and calcium-transient biomarkers as well as the recorded waveforms. In addition, transcriptomic trends were qualitatively compared with the model-inferred conductance changes to assess biological plausibility.

The optimized models reproduced the main experimental features in both conditions and revealed clear condition-specific electrophysiological differences. Both SM and HT remained consistent with an overall immature-like phenotype, characterized by short APD90 values and persistent spontaneous activity. However, HT showed a lower spontaneous beating rate, altered conductances related to repolarization and calcium handling, and greater variability in the inferred parameter space, suggesting a distinct and more heterogeneous electrophysiological state. Partial agreement between transcriptomic and model-inferred trends further supported the biological relevance of several inferred conductance changes.

Overall, this model-informed framework enabled mechanistic characterization of bioengineered hiPSC-CM tissues beyond conventional waveform analysis alone. By linking tissue-level OM recordings with inferred ionic remodeling, this approach may support the evaluation and optimization of maturation strategies in engineered cardiac tissues.