Department of Life Science, National Taiwan University, Taipei 10617, Taiwan.
Genome and Systems Biology Degree Program, National Taiwan University, Taipei 10617, Taiwan.
Cardiovasc Res. 2020 Mar 1;116(3):658-670. doi: 10.1093/cvr/cvz148.
Human embryonic stem cells (hESCs) can be used to generate scalable numbers of cardiomyocytes (CMs) for studying cardiac biology, disease modelling, drug screens, and potentially for regenerative therapies. A fluorescence-based reporter line will significantly enhance our capacities to visualize the derivation, survival, and function of hESC-derived CMs. Our goal was to develop a reporter cell line for real-time monitoring of live hESC-derived CMs.
We used CRISPR/Cas9 to knock a mCherry reporter gene into the MYH6 locus of hESC lines, H1 and H9, enabling real-time monitoring of the generation of CMs. MYH6:mCherry+ cells express atrial or ventricular markers and display a range of cardiomyocyte action potential morphologies. At 20 days of differentiation, MYH6:mCherry+ cells show features characteristic of human CMs and can be used successfully to monitor drug-induced cardiotoxicity and oleic acid-induced cardiac arrhythmia.
We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology.
人类胚胎干细胞（hESC）可用于生成可扩展数量的心肌细胞（CMs），用于研究心脏生物学、疾病建模、药物筛选，以及潜在的再生治疗。荧光报告基因系将极大地增强我们可视化 hESC 衍生的 CMs 的起源、存活和功能的能力。我们的目标是开发一种报告细胞系，用于实时监测活 hESC 衍生的 CMs。
我们使用 CRISPR/Cas9 将 mCherry 报告基因敲入 hESC 系 H1 和 H9 的 MYH6 基因座，从而能够实时监测 CMs 的生成。MYH6:mCherry+细胞表达心房或心室标记物，并显示出多种心肌细胞动作电位形态。在分化 20 天后，MYH6:mCherry+细胞表现出人类 CMs 的特征，可成功用于监测药物诱导的心脏毒性和油酸诱导的心律失常。
我们创建了两个 MYH6:mCherry hESC 报告系，并记录了这些系在与心肌细胞生物学相关的疾病建模中的应用。
