Craig L E, Spelman J P, Strandberg J D, Zink M C
Division of Comparative Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Microvasc Res. 1998 Jan;55(1):65-76. doi: 10.1006/mvre.1997.2045.
An increased recognition of the role of endothelial cells in disease and the development of methods for endothelial cell culture has led to an upsurge in in vitro studies of endothelial cell function. However, the cells most often used for these studies do not reflect the in vivo heterogeneity of endothelial cells. To assess intrinsic differences between large and small vessel endothelial cells from different tissues, primary cultures of endothelial cells from capillaries (brain, lung, and adipose tissue) and a large vessel (aorta) of sheep were isolated, purified by fluorescence-activated cell sorting of acetylated low density lipoprotein (DiI-Ac-LDL) labeled cells, and characterized by phase contrast and ultrastructural morphology, expression of von Willebrand factor, and lack of expression of cytokeratin, smooth muscle actin, and glial fibrillary acidic protein (GFAP). Although all endothelial cells were cultured in the same media, only the brain microvascular endothelial cells demonstrated tight junctions by electron microscopy. Only the large vessel (aortic) endothelial cells contained Weibel-Palade bodies. Expression of von Willebrand factor decreased with passage of cells, but uptake of DiI-Ac-LDL was consistently positive regardless of culture conditions or passage number. These studies demonstrate that the unique ultrastructural characteristics of microvascular and macrovascular endothelial cells are intrinsic to the cells themselves and are not determined by differential culture conditions. This system allows the study of pathologic processes that affect endothelial cells of certain target organs selectively and should more accurately represent the response of tissue-specific endothelial cells in inflammatory processes.
对内皮细胞在疾病中的作用的认识不断提高,以及内皮细胞培养方法的发展,导致了内皮细胞功能体外研究的热潮。然而,这些研究中最常用的细胞并不能反映内皮细胞在体内的异质性。为了评估来自不同组织的大血管和小血管内皮细胞之间的内在差异,分离了绵羊毛细血管(脑、肺和脂肪组织)和一条大血管(主动脉)的内皮细胞原代培养物,通过对乙酰化低密度脂蛋白(DiI-Ac-LDL)标记的细胞进行荧光激活细胞分选进行纯化,并通过相差显微镜和超微结构形态、血管性血友病因子的表达以及细胞角蛋白、平滑肌肌动蛋白和胶质纤维酸性蛋白(GFAP)的不表达进行表征。尽管所有内皮细胞都在相同的培养基中培养,但只有脑微血管内皮细胞通过电子显微镜显示出紧密连接。只有大血管(主动脉)内皮细胞含有Weibel-Palade小体。血管性血友病因子的表达随着细胞传代而降低,但无论培养条件或传代数如何,DiI-Ac-LDL的摄取始终为阳性。这些研究表明,微血管和大血管内皮细胞独特的超微结构特征是细胞本身固有的,而不是由不同的培养条件决定的。该系统允许研究选择性影响某些靶器官内皮细胞的病理过程,并且应该更准确地代表炎症过程中组织特异性内皮细胞的反应。