Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes

Michael Xavier Doss¹ , Johannes Winkler¹ , Shuhua Chen¹ , Rita Hippler-Altenburg¹ , Isaia Sotiriadou¹ , Marcel Halbach¹ , Kurt Pfannkuche¹ , Huamin Liang¹ , Herbert Schulz² , Oliver Hummel² , Norbert Hübner² , Ruth Rottscheidt³ , Jürgen Hescheler¹ and Agapios Sachinidis¹

¹Center of Physiology and Pathophysiology, Institute of Neurophysiology, University of Cologne, Robert Koch Str., 50931 Cologne, Germany

²Max-Delbrueck-Center for Molecular Medicine - MDC, Robert-Rössle Str., 13092 Berlin, Germany

³Institute for Genetics, Department of Evolutionary Genetics, University of Cologne, Zülpicher Str., 50674 Cologne, Germany

author email corresponding author email

Genome Biology 2007, 8:R56doi:10.1186/gb-2007-8-4-r56


Published:	11 April 2007

Subject areas: Cell biology, Development, Genome studies

Abstract

Background

Characterization of gene expression signatures for cardiomyocytes derived from embryonic stem cells will help to define their early biologic processes.

Results

A transgenic α-myosin heavy chain (MHC) embryonic stem cell lineage was generated, exhibiting puromycin resistance and expressing enhanced green fluorescent protein (EGFP) under the control of the α-MHC promoter. A puromycin-resistant, EGFP-positive, α-MHC-positive cardiomyocyte population was isolated with over 92% purity. RNA was isolated after electrophysiological characterization of the cardiomyocytes. Comprehensive transcriptome analysis of α-MHC-positive cardiomyocytes in comparison with undifferentiated α-MHC embryonic stem cells and the control population from 15-day-old embryoid bodies led to identification of 884 upregulated probe sets and 951 downregulated probe sets in α-MHC-positive cardiomyocytes. A subset of upregulated genes encodes cytoskeletal and voltage-dependent channel proteins, and proteins that participate in aerobic energy metabolism. Interestingly, mitosis, apoptosis, and Wnt signaling-associated genes were downregulated in the cardiomyocytes. In contrast, annotations for genes upregulated in the α-MHC-positive cardiomyocytes are enriched for the following Gene Ontology (GO) categories: enzyme-linked receptor protein signaling pathway (GO:0007167), protein kinase activity (GO:0004672), negative regulation of Wnt receptor signaling pathway (GO:0030178), and regulation of cell size (O:0008361). They were also enriched for the Biocarta p38 mitogen-activated protein kinase signaling pathway and Kyoto Encyclopedia of Genes and Genomes (KEGG) calcium signaling pathway.

Conclusion

The specific pattern of gene expression in the cardiomyocytes derived from embryonic stem cells reflects the biologic, physiologic, and functional processes that take place in mature cardiomyocytes. Identification of cardiomyocyte-specific gene expression patterns and signaling pathways will contribute toward elucidating their roles in intact cardiac function.