C16 maturation medium increased cardiomyocyte twitch stress by approximately 5-fold (1.69 vs 0.35 mN mm-2) compared to RPMI + B27 treatment in engineered microtissues.
A novel 16-component culture medium optimized via a differential evolution algorithm significantly advances the physiological, electrophysiological, and metabolic maturation of human iPSC-derived cardiomyocytes.
Absolute Event Rate: 1.69% vs 0.35%
p-value: p=0.0002
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold tremendous promise for in vitro modeling to assess native myocardial function and disease mechanisms, as well as testing drug safety and efficacy. However, current hiPSC-CMs are functionally immature, resembling in vivo CMs of fetal or neonatal developmental states. The use of targeted culture media and organoid formats have been identified as potential high-yield contributors to improve CM maturation. This study presents an hiPSC-CM maturation medium formulation, designed using a differential evolutionary approach targeting metabolic functionality for iterative optimization. Relative to existing high-performing reference formulations, our medium significantly matured morphology, Ca2+ handling, electrophysiology, and metabolism, which was further validated by multi-omic screening, for cells in either pure or co-cultured microtissue formats. Together, these findings not only provide a reliable workflow for highly functional hiPSC-CMs for downstream use, but also demonstrate the power of high-dimensional optimization processes in evoking advanced biological function in vitro.
Callaghan et al. (2026) studied Cardiomyocyte maturation. C16 maturation medium vs. RPMI + B27 medium was evaluated on Twitch stress (mN mm-2) (p=0.0002). C16 maturation medium increased cardiomyocyte twitch stress by approximately 5-fold (1.69 vs 0.35 mN mm-2) compared to RPMI + B27 treatment in engineered microtissues.