Su-Yi Tsai1,2, Zaniar Ghazizadeh3, Hou-Jun Wang1, Sadaf Amin3, Francis A Ortega4, Zohreh Sadat Badieyan3, Zi-Ting Hsu1, Miriam Gordillo3, Ritu Kumar3, David J Christini4,5, Todd Evans3, Shuibing Chen3. 1. Department of Life Science, National Taiwan University, Taipei 10617, Taiwan. 2. Genome and Systems Biology Degree Program, National Taiwan University, Taipei 10617, Taiwan. 3. Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA. 4. Physiology, Biophysics, and Systems Biology Graduate Program, Weill Cornell Medical College, New York, NY 10065, USA. 5. Greenberg Division of Cardiology, Department of Medicine, Weill Cornell Medical College, New York, NY 10065, USA.
Abstract
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 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. CONCLUSION: We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology. Published on behalf of the European Society of Cardiology. All rights reserved.
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 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. CONCLUSION: We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology. Published on behalf of the European Society of Cardiology. All rights reserved.
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