Angiogenic profiling and comparison of immortalized endothelial cells for functional genomics
Angiogenic profiling and comparison of immortalized endothelial cells for functional genomics
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DOI：
10.1016/j.yexcr.2007.08.013
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发表时间：
2008-01-15
影响因子：
3.7
通讯作者：
Griffioen, Arjan W.
中科院分区：
文献类型：
--
作者：
van Beijnum, Judy R.;van der Linden, Edith;Griffioen, Arjan W.
基因组学的努力，在过去的十年中，导致在疾病过程中，包括肿瘤血管生成的推定作用的许多基因的鉴定。为了在功能上验证这些基因，培养的内皮细胞是不可或缺的工具，尽管由于缺乏适当的微环境，这些内皮细胞可能不能完全模拟组织内皮细胞的表型。为了获得代表正常生理学的实验数据，优选使用原代内皮细胞。然而，这些细胞通常在传代次数上受到限制，可能难以获得，并且显示出很大的个体间差异。此外，转染效率在原代细胞中非常有限，阻碍了在功能基因组学和基因功能分析中的应用。在此，我们将永生化内皮细胞- HMEC、RF 24和EVLC 2-与原代HUVEC进行了比较。我们发现，RF 24，并在较小程度上HMEC，类似于主要HUVEC的所有方面检查。与EVLC 2相反，RF 24表达内皮标志物CD 31、CD 34、CD 105、vWF和VE-钙粘蛋白，并且能够在体外迁移和管形成。此外，血管生成生长因子及其受体的表达水平与原代EC相当。此外，尽管原代HUVEC对使用常见亲脂性转染试剂的转染有抵抗力，但HMEC和RF 24可以很容易地转染。因此，这些细胞为血管生成研究中的功能基因组学提供了有价值的工具。(C)2007年爱思唯尔公司All rights reserved.
Genomics efforts of the past decade have resulted in the identification of numerous genes with putative roles in disease processes, including tumor angiogenesis. To functionally validate these genes, cultured endothelial cells are indispensable tools, though these may not completely mimic the phenotype of tissue endothelial cells as the proper microenvironment is lacking. To obtain experimental data representative of normal physiology, the use of primary endothelial cells is preferred. However, these cells are usually limited in passage number, can be difficult to obtain and show great interindividual variety. Furthermore, transfection efficiency is very limited in primary cells, hampering applications in functional genomics and gene function analysis. The use of properly characterized alternative endothelial cell sources is therefore warranted.Here, we compared immortalized endothelial cells - HMEC, RF24 and EVLC2 - with primary HUVEC. We show that RF24, and to a slightly lesser extent HMEC, resembles primary HUVEC most on all facets examined. RF24, in contrast to EVLC2, express the endothelial markers CD31, CD34, CD105, vWF and VE-cadherin, and are capable of migration and tube formation in vitro. Furthermore, the expression levels of angiogenic growth factors and their receptors are comparable to that of primary EC. in addition, whereas primary HUVEC are resistant to transfection using common lipophilic transfection reagents, HMEC and RF24 could be readily transfected. Hence, these cells pose a valuable tool for functional genomics in angiogenesis research. (C) 2007 Elsevier Inc. All rights reserved.