Clinic for Cardiology and Pneumology, University Medical Center Göttingen (UMG), Göttingen, Germany.
DZHK (German Center for Cardiovascular Research), partner site Göttingen, Germany.
JCI Insight. 2018 Jun 21;3(12). doi: 10.1172/jci.insight.99941.
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.
生成源自人诱导多能干细胞（iPSC）的同种型特异性心肌细胞（CM）的均质群体及其全面表型分析对于更好地理解亚型相关疾病机制以及作为开发腔特异性药物的工具至关重要。本研究的目的是应用一种简单有效的方法，在无饲养层条件下将 iPSC 分化为明确的功能性 CM 亚型，并在单细胞和工程心肌（EHM）水平上全面了解心房和心室 iPSC-CM 的分子、细胞生物学和功能特性。通过在基于单层和明确培养的条件下对视黄酸信号的阶段特异性激活，我们表明可以将心脏祖细胞定向为高度同质的心房 CM 群体。通过将 iPSC-CM 亚型的转录组和蛋白质组谱与通过光动作电位和钙成像进行的功能特征以及在 EHM 中的收缩分析相结合，我们证明了心房和心室 iPSC-CM 和 -EHM 分别与心房和心室心肌高度对应。本研究全面了解了心房和心室 iPSC-CM 和 -EHM 的分子和功能特征，并支持它们在疾病建模和腔特异性药物筛选中的适用性。
