title: Phenotypic assays for analyses of pluripotent stem cell-derived cardiomyocytes.
abstract: 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.
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|ICRC, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Pesl
authors.affiliation: CEITEC, Masaryk University, Brno, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Pribyl
authors.affiliation: ICRC, St. Anne's University Hospital, Brno, Czech Republic.|CEITEC, Masaryk University, Brno, Czech Republic.
authors.forename: Guido
authors.initials: G
authors.lastname: Caluori
authors.affiliation: Department of Biomedical Engineering, Faculty of Electrical Engineering and Communication, Brno University of Technology, Brno, Czech Republic.
authors.forename: Vratislav
authors.initials: V
authors.lastname: Cmiel
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Ivana
authors.initials: I
authors.lastname: Acimovic
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Sarka
authors.initials: S
authors.lastname: Jelinkova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|ICRC, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Dvorak
authors.affiliation: ICRC, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Starek
authors.affiliation: CEITEC, Masaryk University, Brno, Czech Republic.|Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Skladal
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|ICRC, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Rotrekl
mesh_terms: D002454:Cell Differentiation; D005838:Genotype; D057166:High-Throughput Screening Assays; D006801:Humans; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000166:cytology*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology*
keywords: atomic force microscopy; cardiomyocytes; differentiation; drug screening; drug toxicity; hESC; hiPSC; high-throughput screening