Institute of Experimental and Clinical Pharmacology and Toxicology, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
Heart Center Freiburg University, Department of Cardiology and Angiology I, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
Sci Rep. 2020 Feb 10;10(1):2241. doi: 10.1038/s41598-020-59213-x.
Endothelial cells take pivotal roles in the heart and the vascular system and their differentiation, subspecification and function is determined by gene expression. A stable, in vitro cardiac endothelial cell line could provide high cell numbers as needed for many epigenetic analyses and facilitate the understanding of molecular mechanisms involved in endothelial cell biology. To test their suitability for transcriptomic or epigenetic studies, we compared the transcriptome of cultured immortalized mouse cardiac endothelial cells (MCEC) to primary cardiac endothelial cells (pEC). Whole transcriptome comparison of MCEC and pEC showed a correlation of 0.75-0.77. Interestingly, correlation of gene expression declined in endothelial cell-typical genes. In MCEC, we found a broad downregulation of genes that are highly expressed in pEC, including well-described markers of endothelial cell differentiation. Accordingly, systematic analysis revealed a downregulation of genes associated with typical endothelial cell functions in MCEC, while genes related to mitotic cell cycle were upregulated when compared to pEC. In conclusion, the findings from this study suggest that primary cardiac endothelial cells should preferably be used for genome-wide transcriptome or epigenome studies. The suitability of in vitro cell lines for experiments investigating single genes or signaling pathways should be carefully validated before use.
内皮细胞在心脏和血管系统中起着至关重要的作用,其分化、亚特化和功能取决于基因表达。稳定的体外心脏内皮细胞系可以提供大量所需的细胞,用于许多表观遗传学分析,并有助于理解内皮细胞生物学中涉及的分子机制。为了测试它们是否适合转录组学或表观遗传学研究,我们比较了培养的永生化小鼠心脏内皮细胞 (MCEC) 和原代心脏内皮细胞 (pEC) 的转录组。MCEC 和 pEC 的全转录组比较显示相关性为 0.75-0.77。有趣的是,内皮细胞典型基因的表达相关性下降。在 MCEC 中,我们发现高表达于 pEC 的基因广泛下调,包括已描述的内皮细胞分化标志物。因此,系统分析显示,与 pEC 相比,MCEC 中与典型内皮细胞功能相关的基因下调,而与有丝分裂细胞周期相关的基因上调。总之,这项研究的结果表明,原代心脏内皮细胞应优先用于全基因组转录组或表观基因组研究。在使用之前,应仔细验证体外细胞系用于研究单个基因或信号通路的实验的适用性。