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Directed differentiation of human embryonic stem cells to interrogate the cardiac gene regulatory network.

Authors :
Dixon JE
Dick E
Rajamohan D
Shakesheff KM
Denning C
Source :
Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2011 Sep; Vol. 19 (9), pp. 1695-703. Date of Electronic Publication: 2011 Jun 21.
Publication Year :
2011

Abstract

The limited ability of the heart to regenerate has prompted development of new systems to produce cardiomyocytes for therapeutics. While differentiation of human embryonic stem cells (hESCs) into cardiomyocytes has been well documented, the process remains inefficient and/or expensive, and progress would be facilitated by better understanding the early genetic events that cause cardiac specification. By maintaining a transgenic cardiac-specific MYH6-monomeric red fluorescent protein (mRFP) reporter hESC line in conditions that promote pluripotency, we tested the ability of combinations of 15 genes to induce cardiac specification. Screening identified GATA4 plus TBX5 as the minimum requirement to activate the cardiac gene regulatory network and produce mRFP(+) cells, while a combination of GATA4, TBX5, NKX2.5, and BAF60c (GTNB) was necessary to generate beating cardiomyocytes positive for cTnI and α-actinin. Including the chemotherapeutic agent, Ara-C, from day 10 of induced differentiation enriched for cTnI/α-actinin double positive cells to 45%. Transient expression of GTNB for 5-7 days was necessary to activate the cardiogenesis through progenitor intermediates in a manner consistent with normal heart development. This system provides a route to test the effect of different factors on human cardiac differentiation and will be useful in understanding the network failures that underlie disease phenotypes.

Details

Language :
English
ISSN :
1525-0024
Volume :
19
Issue :
9
Database :
MEDLINE
Journal :
Molecular therapy : the journal of the American Society of Gene Therapy
Publication Type :
Academic Journal
Accession number :
21694703
Full Text :
https://doi.org/10.1038/mt.2011.125