A COUP-TFII Human Embryonic Stem Cell Reporter Line to Identify and Select Atrial Cardiomyocytes

Autor: Verena Schwach, Mervyn P.H. Mol, Valeria V. Orlova, Robert Passier, Konstantinos Anastassiadis, Arie O. Verkerk, Richard P. Davis, Christine L. Mummery, Jantine Monshouwer-Kloots, Harsha D. Devalla
Přispěvatelé: ACS - Amsterdam Cardiovascular Sciences, Medical Biology, Cardiology, Applied Stem Cell Technology
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Pluripotent Stem Cells
Ovalbumin
Cellular differentiation
Green Fluorescent Proteins
cardiac differentiation
Chick Embryo
Biology
Biochemistry
Article
Green fluorescent protein
COUP Transcription Factor II
03 medical and health sciences
Mice
Genes
Reporter
Genetics
Animals
Humans
Myocytes
Cardiac
Heart Atria
cardiovascular diseases
Progenitor cell
Induced pluripotent stem cell
Promoter Regions
Genetic
lcsh:QH301-705.5
CRISPR/Cas9
COUP-TFII
lcsh:R5-920
Cell Differentiation
Cell Biology
human embryonic stem cells
Molecular biology
Embryonic stem cell
COUP-TFII-mCherry fluorescent stem cell reporter
3. Good health
030104 developmental biology
lcsh:Biology (General)
Cell culture
Cell Tracking
COUP-TFII-knockout
cardiovascular system
CRISPR-Cas Systems
lcsh:Medicine (General)
mCherry
Developmental Biology
atrial specification
Zdroj: Stem Cell Reports
Stem Cell Reports, Vol 9, Iss 6, Pp 1765-1779 (2017)
Stem Cell Reports, 9(6), 1765-1779
Stem cell reports, 9(6), 1765-1779. Cell Press
ISSN: 2213-6711
Popis: Summary Reporter cell lines have already proven valuable in identifying, tracking, and purifying cardiac subtypes and progenitors during differentiation of human pluripotent stem cells (hPSCs). We previously showed that chick ovalbumin upstream promoter transcription factor II (COUP-TFII) is highly enriched in human atrial cardiomyocytes (CMs), but not ventricular. Here, we targeted mCherry to the COUP-TFII genomic locus in hPSCs expressing GFP from the NKX2.5 locus. This dual atrial NKX2.5EGFP/+-COUP-TFIImCherry/+ reporter line allowed identification and selection of GFP+ (G+)/mCherry+ (M+) CMs following cardiac differentiation. These cells exhibited transcriptional and functional properties of atrial CMs, whereas G+/M− CMs displayed ventricular characteristics. Via CRISPR/Cas9-mediated knockout, we demonstrated that COUP-TFII is not required for atrial specification in hPSCs. This new tool allowed selection of human atrial and ventricular CMs from mixed populations, of relevance for studying cardiac specification, developing human atrial disease models, and examining distinct effects of drugs on the atrium versus ventricle.
Graphical Abstract
Highlights • Dual NKX2.5EGFP/+-COUP-TFIImCherry/+ hPSCs to identify atrial cardiomyocytes • COUP-TFII is not required for atrial specification of hPSCs in vitro
In this article, Passier and colleagues developed an atrial fluorescent stem cell reporter by CRISPR/Cas9-mediated knockin of mCherry at the genomic locus of COUP-TFII in a cardiac NKX2.5EGFP/w reporter line. As validated at the transcriptional and functional level, the nuclear receptor COUP-TFII successfully marks atrial cardiomyocytes but is not required for atrial specification during in vitro differentiation of hPSCs.
Databáze: OpenAIRE
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