alexa Dgcr8 is Indispensable for Cardiac Lineage Specification in Embryonic Stem Cells
ISSN: 2157-7633

Journal of Stem Cell Research & Therapy
Open Access

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Research Article

Dgcr8 is Indispensable for Cardiac Lineage Specification in Embryonic Stem Cells

Zhiyong Lei1, Alain van Mil1, Annebel M van de Vrugt1, Pieter A Doevendans1,2 and Joost PG Sluijter1,2*

1Department of Cardiology, Division Heart and Lungs, University Medical Center Utrecht, Utrecht, The Netherlands

2ICIN, Netherlands Heart Institute, Moreelsepark 1, 3511 EP, Utrecht, The Netherlands

*Corresponding Author:
Joost PG Sluijter
Division of Heart and Lungs
Department of Cardiology, Experimental Cardiology Laboratory
Huispostnummer G02.523, Postbus 85500
3508 GA Utrecht, The Netherlands,
Tel: + 31 88 755 7567
Fax: + 31 30 252 2693
E-mail: [email protected]

Received date: December 15, 2014; Accepted date: January 13, 2015; Published date: January 15, 2015

Citation: Lei Z, van Mil A, van de Vrugt AM, Doevendans PA, Sluijter JPG (2015) Dgcr8 is Indispensable for Cardiac Lineage Specification in Embryonic Stem Cells. J Stem Cell Res Ther 5:260. doi: 10.4172/2157-7633.1000260

Copyright: ©2015 Lei Z, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Objective: microRNAs have been shown to play important roles in cellular behavior and lineage specification including cardiogenic differentiation. However, full understanding of their roles in cardiomyocyte differentiation has been impeded due to lack of proper cellular model. Here, we used an embryonic stem cell (ESC) that is lacking the important microprocessor Dgcr8 (or Pasha), which allows the introduction of individual miRNAs to study their role in cardiac differentiation and for more precise target selection.
Methods: Dgcr8 KO ESC was cultured in LIF-supplemented ESC medium with mouse embryonic fibroblast feeders and cardiac differentiation was induced using an embryonic body-based differentiation protocol. Differentiation was monitored by measuring mRNA and protein levels of cardiogenic markers and heterochromatin changes using immunofluorescent staining and semi-quantitative PCR.
Results and conclusion: We showed that Dgcr8 KO ESCs indeed are lacking a large population of small RNAs, including but not limited to mature microRNAs. The KO cells had a lower proliferation rate and were unable to differentiate into the cardiac lineage. To our surprise, in addition to a defect in microRNA processing, Dgcr8 KO embryonic stem cells are unable to form proper heterochromatin and to inactivate genotoxic centromeric repetitive elements. Our results argue that, in addition to controlling microRNA processing, Dgcr8 may serve a previously unrecognized role in heterochromatin silencing.

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