JCI Insight. 2018 Jun 21;3(12). pii: 99941. doi: 10.1172/jci.insight.99941. [Epub ahead of print]

Deep phenotyping of human induced pluripotent stem cell-derived atrial and ventricular cardiomyocytes.

Cyganek L^1,2, Tiburcy M^2,3, Sekeres K⁴, Gerstenberg K¹, Bohnenberger H⁵, Lenz C^6,7, Henze S¹, Stauske M^1,2, Salinas G⁸, Zimmermann WH^2,3, Hasenfuss G^1,2, Guan K^1,2,4.

Abstract

Generation of homogeneous populations of subtype-specific cardiomyocytes (CMs) derived from human induced pluripotent stem cells (iPSCs) and their comprehensive phenotyping is crucial for a better understanding of the subtype-related disease mechanisms and as tools for the development of chamber-specific drugs. The goals of this study were to apply a simple and efficient method for differentiation of iPSCs into defined functional CM subtypes in feeder-free conditions and to obtain a comprehensive understanding of the molecular, cell biological, and functional properties of atrial and ventricular iPSC-CMs on both the single-cell and engineered heart muscle (EHM) level. By a stage-specific activation of retinoic acid signaling in monolayer-based and well-defined culture, we showed that cardiac progenitors can be directed towards a highly homogeneous population of atrial CMs. By combining the transcriptome and proteome profiling of the iPSC-CM subtypes with functional characterizations via optical action potential and calcium imaging, and with contractile analyses in EHM, we demonstrated that atrial and ventricular iPSC-CMs and -EHM highly correspond to the atrial and ventricular heart muscle, respectively. This study provides a comprehensive understanding of the molecular and functional identities characteristic of atrial and ventricular iPSC-CMs and -EHM and supports their suitability in disease modeling and chamber-specific drug screening.