Geometry-dependent functional changes in iPSC-derived cardiomyocytes probed by functional imaging and RNA sequencing
Christopher A. Werley,Miao-Ping Chien,Jellert Gaublomme,Karthik Shekhar,Vincent L. Butty,B. Alexander Yi,Joel M. Kralj,William Bloxham,Laurie A. Boyer,Aviv Regev,Aviv Regev,Aviv Regev,Adam E. Cohen,Adam E. Cohen +13 more
TL;DR: The data show that optical electrophysiology is a powerful tool for assaying hiPSC-CM maturation, and that island size powerfully drives activation of a subset of genes involved in cardiac maturation.
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Abstract: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are a promising platform for cardiac studies in vitro, and possibly for tissue repair in humans. However, hiPSC-CM cells tend to retain morphology, metabolism, patterns of gene expression, and electrophysiology similar to that of embryonic cardiomyocytes. We grew hiPSC-CM in patterned islands of different sizes and shapes, and measured the effect of island geometry on action potential waveform and calcium dynamics using optical recordings of voltage and calcium from 970 islands of different sizes. hiPSC-CM in larger islands showed electrical and calcium dynamics indicative of greater functional maturity. We then compared transcriptional signatures of the small and large islands against a developmental time course of cardiac differentiation. Although island size had little effect on expression of most genes whose levels differed between hiPSC-CM and adult primary CM, we identified a subset of genes for which island size drove the majority (58%) of the changes associated with functional maturation. Finally, we patterned hiPSC-CM on islands with a variety of shapes to probe the relative contributions of soluble factors, electrical coupling, and direct cell-cell contacts to the functional maturation. Collectively, our data show that optical electrophysiology is a powerful tool for assaying hiPSC-CM maturation, and that island size powerfully drives activation of a subset of genes involved in cardiac maturation.
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Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine
Elaheh Karbassi,Aidan M. Fenix,Silvia Marchianò,Naoto Muraoka,Kenta Nakamura,Xiulan Yang,Charles E. Murry +6 more
TL;DR: The hallmarks of cardiomyocyte maturation are described and the current approaches to mature stem cell-derivedCardiomyocytes with adult-like phenotypes in vitro are presented, highlighting challenges and future directions to generate cardiomers with an optimal maturation state for use in research and regenerative medicine.
All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins
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