| Title | A human embryonic stem cell reporter line for monitoring chemical-induced cardiotoxicity. |
| Publication Type | Journal Article |
| Year of Publication | 2020 |
| Authors | Tsai S-Y, Ghazizadeh Z, Wang H-J, Amin S, Ortega FA, Badieyan ZSadat, Hsu Z-T, Gordillo M, Kumar R, Christini DJ, Evans T, Chen S |
| Journal | Cardiovasc Res |
| Volume | 116 |
| Issue | 3 |
| Pagination | 658-670 |
| Date Published | 2020 Mar 01 |
| ISSN | 1755-3245 |
| Keywords | 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, Red Fluorescent Protein, Time Factors |
| Abstract | <p><b>AIMS: </b>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.</p><p><b>METHODS AND RESULTS: </b>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.</p><p><b>CONCLUSION: </b>We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology.</p> |
| DOI | 10.1093/cvr/cvz148 |
| Alternate Journal | Cardiovasc Res |
| PubMed ID | 31173076 |
| PubMed Central ID | PMC7252441 |
| Grant List | R35 HL135778 / HL / NHLBI NIH HHS / United States |