Phenotypic assays for analyses of pluripotent stem cell-derived cardiomyocytes.
J Mol Recognit. 2016-12-20; 30(6): e2602
DOI: 10.1002/jmr.2602
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Pesl M(1)(2), Pribyl J(3), Caluori G(2)(3), Cmiel V(4), Acimovic I(1), Jelinkova S(1), Dvorak P(1)(2), Starek Z(2), Skladal P(3)(5), Rotrekl V(1)(2).
Author information:
(1)Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
(2)ICRC, St. Anne’s University Hospital, Brno, Czech Republic.
(3)CEITEC, Masaryk University, Brno, Czech Republic.
(4)Department of Biomedical Engineering, Faculty of Electrical Engineering and Communication, Brno University of Technology, Brno, Czech Republic.
(5)Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
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.
Copyright © 2016 John Wiley & Sons, Ltd.