Synnergren Jane, Akesson Karolina, Dahlenborg Kerstin, Vidarsson Hilmar, Améen Caroline, Steel Daniella, Lindahl Anders, Olsson Björn, Sartipy Peter
School of Humanities and Informatics, University of Skövde, P.O. Box 408, SE-541 28 Skövde, Sweden.
Stem Cells. 2008 Jul;26(7):1831-40. doi: 10.1634/stemcells.2007-1033. Epub 2008 Apr 24.
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. Real-time 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.
人类胚胎干细胞（hESCs）可在体外分化为自发收缩的心肌细胞（CMs）。这些细胞在基础研究、药物发现和再生医学的各种应用中可能极具价值。为了充分利用这些细胞的潜力，需要对它们进行广泛的表征，并且需要更好地确定控制hESC向心脏谱系分化的调控机制。在本研究中，我们首次使用微阵列分析分离的hESC来源的CM簇的全局基因表达谱。通过将这些簇与未分化的hESCs进行比较并使用严格的选择标准，我们在收缩簇中鉴定出530个上调基因和40个下调基因。为了进一步表征hESC来源的CM簇中上调基因家族，根据其基因本体注释对这些基因进行分类。结果表明，hESC来源的CM簇在分子水平上与人类心脏组织具有高度相似性。此外，利用上调基因家族，我们创建了揭示拓扑特征的蛋白质相互作用图谱。我们还在hESC来源的CM簇中的上调基因中搜索细胞途径，并鉴定出八条显著上调的途径。实时定量聚合酶链反应和免疫组织化学分析证实了微阵列鉴定的一部分基因的表达。综上所述，本文呈现的结果提供了hESC来源的CM簇的分子特征，并进一步加深了我们对这些细胞中活跃的生物学过程的理解。
