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Phenotypic assays for analyses of pluripotent stem cell-derived cardiomyocytes

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PEŠL Martin PŘIBYL Jan CALUORI Guido CMIEL Vratislav AĆIMOVIĆ Ivana JELÍNKOVÁ Šárka DVOŘÁK Petr STÁREK Zdeněk SKLÁDAL Petr ROTREKL Vladimír

Druh Článek v odborném periodiku
Časopis / Zdroj Journal of Molecular Recognition
Fakulta / Pracoviště MU

Lékařská fakulta Přírodovědecká fakulta Středoevropský technologický institut

Citace
WWW http://dx.doi.org/10.1002/jmr.2602
Doi http://dx.doi.org/10.1002/jmr.2602
Obor Genetika a molekulární biologie
Klíčová slova Atomic force microscopy; Cardiomyocytes; Differentiation; Drug screening; Drug toxicity; HESC; High-throughput screening; HiPSC
Popis Stem cell-derived cardiomyocytes (CMs) hold great hopes for myocardium regeneration because of their ability to produce functional cardiac cells in large quantities. They also hold promise in dissecting the molecular principles involved in heart diseases and also in drug development, owing to their ability to model the diseases using patient-specific human pluripotent stem cell (hPSC)-derived CMs. The CM properties essential for the desired applications are frequently evaluated through morphologic and genotypic screenings. Even though these characterizations are necessary, they cannot in principle guarantee the CM functionality and their drug response. The CM functional characteristics can be quantified by phenotype assays, including electrophysiological, optical, and/or mechanical approaches implemented in the past decades, especially when used to investigate responses of the CMs to known stimuli (eg, adrenergic stimulation). Such methods can be used to indirectly determine the electrochemomechanics of the cardiac excitation-contraction coupling, which determines important functional properties of the hPSC-derived CMs, such as their differentiation efficacy, their maturation level, and their functionality. In this work, we aim to systematically review the techniques and methodologies implemented in the phenotype characterization of hPSC-derived CMs. Further, we introduce a novel approach combining atomic force microscopy, fluorescent microscopy, and external electrophysiology through microelectrode arrays. We demonstrate that this novel method can be used to gain unique information on the complex excitation-contraction coupling dynamics of the hPSC-derived CMs.
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