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.

A Brief Review of Current Maturation Methods for Human Induced Pluripotent Stem Cells-Derived Cardiomyocytes

“…In hiPSC-CMs, T-tubules are absent and SR is underdeveloped with low expression of SERCA and other key proteins. As a result, hiPSC-CMs rely on L-type channels for the increase of Ca 2+ and ECC is slow (Pesl et al, 2017;Veerman et al, 2017).…”

Fifteen years of Servitude et Grandeur to the application of a biophysical technique in medicine: The tale of AFMBioMed

“…A virtual issue appeared in the Journal of Molecular Recognition in 2017 . Nine research articles composed this special issue . At the end of the conference, it was announced that the next one will be organized in Kraków, in 2017.…”

“…[48][49][50][51][52][53][54][55][56][57] The last change that was made during the 75 Nine research articles composed this special issue. [76][77][78][79][80][81][82][83][84] At the end of the conference, it was announced that the next one will…”

AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

“…Recently, several groups have characterized the mechanical properties of CMs using the force sensitivity of the Atomic Force Microscope (AFM), and its capability of working in physiological environment. The AFM has been used to directly measure the strength of cardiomyocytes at the single cell level, in small clusters or even in large beating clusters (beating bodies—BBs) . Due to its peculiarities, AFM is one of the few techniques that can directly access beating rate and intensity, from an entire BB under physiological conditions and under the action of several drugs.…”

“…The AFM has been used to directly measure the strength of cardiomyocytes at the single cell level, [16][17][18] in small clusters 19 or even in large beating clusters (beating bodies-BBs). 20 Due to its peculiarities, AFM is one of the few techniques that can directly access beating rate and intensity, from an entire BB under physiological conditions and under the action of several drugs. For instance, a recent work 21 from Rotrekl's group have shown how AFM can monitor WT BBs under mechanical stress conditions, while at the same time recording the cellular calcium and electrophysiology response.…”