A Human Embryonic Stem Cell Reporter Line for Monitoring Chemical-induced Cardiotoxicity
Journal
Cardiovascular Research
Journal Volume
116
Journal Issue
3
Pages
658
Date Issued
2020
Author(s)
Tsai, Su-Yi
Ghazizadeh, Zaniar
Wang, Hou-Jun
Ortega, Francis A.
Badieyan, Zohreh S.
Hsu, Zi-Ting
Gordillo, Miriam
Kumar, Ritu
Christini, David J.
Evans, Todd
Chen, Shuibing
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. ? 2019 Published on behalf of the European Society of Cardiology. All rights reserved.
Subjects
Cardiomyocyte; Cardiotoxicity testing; Disease model; hESC reporter; MYH6
SDGs
Other Subjects
actc1 protein; alpha actinin 2; bax2 protein; biological marker; caspase 3; CD34 antigen; doxorubicin; genomic DNA; myh6 protein; myl2 protein; myl7 protein; oleic acid; platelet endothelial cell adhesion molecule 1; Thy 1 membrane glycoprotein; tnni3 protein; tnnt2 protein; transcription factor GATA 4; transcriptome; unclassified drug; vascular cell adhesion molecule 1; biological marker; cardiac myosin; doxorubicin; MYH6 protein, human; myosin heavy chain; oleic acid; photoprotein; red fluorescent protein; action potential; adult; APD90; apoptosis; Article; calcium cell level; cardiac muscle cell; cardiotoxicity; cell differentiation; cell population; controlled study; CRISPR-CAS9 system; DNA sequencing; gene expression profiling; gene targeting; heart arrhythmia; human; human cell; human embryonic stem cell; priority journal; promoter region; reporter gene; RNA sequencing; stem cell culture; stop codon; whole cell patch clamp; biosynthesis; cardiac muscle cell; cardiotoxicity; cell differentiation; cell line; CRISPR Cas system; drug effect; gene knock-in; genetics; heart disease; human embryonic stem cell; metabolism; pathology; pathophysiology; time factor; Action Potentials; Arrhythmias, Cardiac; Biomarkers; Cardiac Myosins; Cardiotoxicity; Cell Differentiation; Cell Line; CRISPR-Cas Systems; Doxorubicin; Gene Knock-In Techniques; Genes, Reporter; Heart Diseases; Human Embryonic Stem Cells; Humans; Luminescent Proteins; Myocytes, Cardiac; Myosin Heavy Chains; Oleic Acid; Time Factors
Type
journal article