期刊
CARDIOVASCULAR RESEARCH
卷 116, 期 3, 页码 658-670出版社
OXFORD UNIV PRESS
DOI: 10.1093/cvr/cvz148
关键词
Cardiomyocyte; Cardiotoxicity testing; MYH6; hESC reporter; Disease model
资金
- Empire State Stem Cell Research Program [NYSTEM] [C028115]
- National Institutes of Health [R35-HL135778]
- American Heart Association [18CSA34080171]
- Ministry of Science and Technology, Taiwan [MOST 105-2320-B-002-065-MY2, 107-2320-B-002026]
Aims 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. Methods and results 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 20days 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. Conclusion We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology.
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