期刊
CARDIOVASCULAR RESEARCH
卷 118, 期 3, 页码 828-843出版社
OXFORD UNIV PRESS
DOI: 10.1093/cvr/cvab102
关键词
hESCs; Heart field-specific cardiomyocytes; Cardiac differentiation; Corin; Electrophysiology
资金
- Eli & Edythe Broad Center of Regenerative Medicine and Stem Cell Research at UCLA Postdoctoral Fellowship
- Department of Defense [PR182456]
- Ruth L. Kirschstein Predoctoral Fellowship [HL144057]
- California Institute for Regenerative Medicine (CIRM) [RN3-06378]
- National Institute of Health [R01HL148714]
- UCLA BSCRC-Rose Hills Foundation Research Award
- Eli and Edythe Broad Foundation Innovative Pilot Stem Cell Research Grant
In this study, we successfully isolated and characterized first and second heart field- and nodal-like cardiomyocytes from human embryonic stem cells using a double reporter line. Our findings provide a comprehensive understanding of the early development and functional properties of these different cardiomyocyte populations. We also identified two novel surface markers that can be used for purification of cardiomyocytes. This research has important implications for gene expression profiling, arrhythmia modeling, and drug screening, and it paves the way for the development of stem cell therapy for specific heart conditions.
Aims We prospectively isolate and characterize first and second heart field- and nodal-like cardiomyocytes using a double reporter line from human embryonic stem cells. Our double reporter line utilizes two important transcription factors in cardiac development, TBX5 and NKX2-5. TBX5 expression marks first heart field progenitors and cardiomyocytes while NKX2-5 is expressed in nearly all myocytes of the developing heart (excluding nodal cells). We address the shortcomings of prior work in the generation of heart field-specific cardiomyocytes from induced pluripotent stem cells and provide a comprehensive early developmental transcriptomic as well as electrophysiological analyses of these three populations. Methods and results Transcriptional, immunocytochemical, and functional studies support the cellular identities of isolated populations based on the expression pattern of NKX2-5 and TBX5. Importantly, bulk and single-cell RNA sequencing analyses provide evidence of unique molecular signatures of isolated first and second heart field cardiomyocytes, as well as nodal-like cells. Extensive electrophysiological analyses reveal dominant atrial action potential phenotypes in first and second heart fields in alignment with our findings in single-cell RNA sequencing. Lastly, we identify two novel surface markers, POPDC2 and CORIN, that enable purification of cardiomyocytes and first heart field cardiomyocytes, respectively. Conclusions We describe a high-yield approach for isolation and characterization of human embryonic stem cell-derived heart field-specific and nodal-like cardiomyocytes. Obtaining enriched populations of these different cardiomyocyte subtypes increases the resolution of gene expression profiling during early cardiogenesis, arrhythmia modelling, and drug screening. This paves the way for the development of effective stem cell therapy to treat diseases that affect specific regions of the heart- or chamber-specific congenital heart defects.
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