Advanced physiological maturation of iPSC-derived human cardiomyocytes using an algorithm-directed optimization of defined media components

Callaghan, N. I.; Durland, L. J.; Chen, W.; Kuzmanov, U.; Miranda, M. Z.; Mirzaei, Z.; Ireland, R. G.; Wang, E. Y.; Wagner, K.; Kim, M. M.; Audet, J.; Santerre, P.; Gramolini, A. O.; Billia, F.; Radisic, M.; Ellis, J.; Mital, S.; Backx, P. H.; Simmons, C. A.

doi:10.1101/2022.10.10.507929

bioRxiv · 10.1101/2022.10.10.507929

Advanced physiological maturation of iPSC-derived human cardiomyocytes using an algorithm-directed optimization of defined media components

Callaghan, N. I.·Durland, L. J.·Chen, W.·Kuzmanov, U.·Miranda, M. Z.·Mirzaei, Z.·Ireland, R. G.·Wang, E. Y.·Wagner, K.·Kim, M. M.·Audet, J.·Santerre, P.·Gramolini, A. O.·Billia, F.·Radisic, M.·Ellis, J.·Mital, S.·Backx, P. H.·Simmons, C. A.

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Abstract

Induced pluripotent stem cell-derived cardiomyocytes (iPSC-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 iPSC- 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 a novel iPSC-CM maturation medium formulation, designed using a differential evolutionary approach targeting metabolic functionality for iterative optimization. Relative to gold-standard reference formulations, our medium significantly matured morphology, Ca2+ handling, electrophysiology, and metabolism, which was further validated by multiomic screening, for cells in either pure or co-cultured microtissue formats. Together, these findings not only provide a reliable workflow for highly functional iPSC-CMs for downstream use, but also demonstrate the power of high-dimensional optimization processes in evoking advanced biological function in vitro.

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Callaghan, N. I., Durland, L. J., Chen, W., Kuzmanov, U., Miranda, M. Z., Mirzaei, Z., Ireland, R. G., Wang, E. Y., Wagner, K., Kim, M. M., Audet, J., Santerre, P., Gramolini, A. O., Billia, F., Radisic, M., Ellis, J., Mital, S., Backx, P. H., Simmons, C. A.. 2022-10-13. Advanced physiological maturation of iPSC-derived human cardiomyocytes using an algorithm-directed optimization of defined media components. https://doi.org/10.1101/2022.10.10.507929

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