Molecular Signature of Cardiomyocyte Clusters Derived from Human Embryonic Stem Cells

Abstract

Human embryonic stem cells (hESCs) can differentiate in vitro into spontaneously contracting cardiomyocytes (CMs). These cells may prove extremely useful for various applications in basic research, drug discovery, and regenerative medicine. To fully use the potential of the cells, they need to be extensively characterized, and the regulatory mechanisms that control hESC differentiation toward the cardiac lineage need to be better defined. In this study, we used microarrays to analyze, for the first time, the global gene expression profile of isolated hESC-derived CM clusters. By comparing the clusters with undifferentiated hESCs and using stringent selection criteria, we identified 530 upregulated and 40 downregulated genes in the contracting clusters. To further characterize the family of upregulated genes in the hESC-derived CM clusters, the genes were classified according to their Gene Ontology annotation. The results indicate that the hESC-derived CM clusters display high similarities, on a molecular level, to human heart tissue. Moreover, using the family of upregulated genes, we created protein interaction maps that revealed topological characteristics. We also searched for cellular pathways among the upregulated genes in the hESC-derived CM clusters and identified eight significantly upregulated pathways. Realtime quantitative polymerase chain reaction and immunohistochemical analysis confirmed the expression of a subset of the genes identified by the microarrays. Taken together, the results presented here provide a molecular signature of hESC-derived CM clusters and further our understanding of the biological processes that are active in these cells.

Comparison of human cardiac gene expression profiles in paired samples of right atrium and left ventricle collected in vivo

“…To investigate possible interactions between the gene products from more abundantly expressed genes in RA and LV samples, the STRING search tool (25,30) was used for creation of protein interaction networks (PIN) as previously described (27). To increase the validity of our results, this search was restricted to include only information extracted from biological experiments and confident interaction databases (30).…”

Global DNA methylation and transcriptional analyses of human ESC-derived cardiomyocytes

“…Recent efforts have successfully developed several protocols, by which hESCs can be differentiated into hCMs (Yang et al, 2008;Kattman et al, 2011;Lian et al, 2012;Willems et al, 2012;Zhang et al, 2012). Furthermore, hESC-derived hCMs have been analyzed by microarray and a group of cardiospecific genes were revealed (Beqqali et al, 2006;Cao et al, 2008;Synnergren et al, 2008). However, one inevitable concern is that by these published differentiation protocols, it is difficult to obtain pure hCMs.…”

Differentiation of Human Atrial Myocytes From Endothelial Progenitor Cell-Derived Induced Pluripotent Stem Cells

“…An alternative method for cardiac differentiation is based on the role endoderm cells play to provide signals to adjacent mesoderm cell for during embryonic development [227][228][229][230]. Visceral endoderm like cells (END2), which derive from mouse P19 embryonal carcinoma, have been designed for cardiac induction.…”

“…Cardiac differentiation from pluripotent stem cells can be derived by guided differentiation in adherent culture involving serum free medium supplemented with growth factors Activin A and BMP4 [55,[221][222][223][224][225][226][227][228][229][230][231][232][233][234][235][236][237][238][239]. These growth factors are a member of the TGF b family that plays roles in embryonic development including induction of gastrulation-like events, meso-endoderm development and induction of cardiogenesis [240][241][242][243][244][245][246].…”