Nitta K, Uchida K, Yumura W, Nihei H
Department of Medicine, Kidney Center, Tokyo Women's Medical College, Japan.
Nihon Jinzo Gakkai Shi. 1993 Aug;35(8):887-91.
Because of difficulties associated with the culture, cloning and propagation of glomerular endothelial cells (GENs), the biological properties of these cells remain largely unknown. We modified the methods established by Ballermann to propagate GENs from adult bovine kidney. We found that the addition of insulin, transferrin and selenium into the standard culture media was an important step in promoting the propagation of the first clone from a single cell and in maintaining the viability of the cells. These cells expressed factor VIII-related antigen and took up acetylated-LDL, but did not contain the Weibel-Palade body, unlike endothelial cells derived from large vessels. Furthermore, GENs were compared with aortic endothelial cells (AECs) to investigate the differences in culture conditions. Compared with AECs, GENs required a higher concentration of serum and the supplementation of growth factor to maintain their biological activity. In addition, GENs were very susceptible to trypsinization and produced prostaglandin E2 as a major cyclooxygenase product, whereas AECs produced PGI2. These findings suggest that GENs will be easily obtained from adult bovine kidney in culture and provide useful information on the functional properties of these cells under physiological and pathophysiological conditions.
由于肾小球内皮细胞(GENs)的培养、克隆和增殖存在困难,这些细胞的生物学特性在很大程度上仍不为人知。我们改进了Ballermann建立的从成年牛肾脏中增殖GENs的方法。我们发现,在标准培养基中添加胰岛素、转铁蛋白和硒是促进从单个细胞培养出第一个克隆以及维持细胞活力的重要步骤。这些细胞表达VIII因子相关抗原并摄取乙酰化低密度脂蛋白,但与大血管来源的内皮细胞不同,它们不含魏-帕小体。此外,将GENs与主动脉内皮细胞(AECs)进行比较以研究培养条件的差异。与AECs相比,GENs需要更高浓度的血清并补充生长因子以维持其生物学活性。此外,GENs对胰蛋白酶消化非常敏感,主要产生前列腺素E2作为环氧化酶产物,而AECs产生前列环素(PGI2)。这些发现表明,GENs在培养中很容易从成年牛肾脏获得,并为这些细胞在生理和病理生理条件下的功能特性提供有用信息。