Carley W W, Milici A J, Madri J A
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.
Exp Cell Res. 1988 Oct;178(2):426-34. doi: 10.1016/0014-4827(88)90411-9.
Capillary endothelial cells in a fenestrated vasculature contain openings in attenuated areas of the cytoplasm which are often covered with one or two diaphragms. However, due to endothelial cell dedifferentiation upon culturing, such indicative membrane structure are often not expressed. The expression of a more differentiated phenotype can be regulated by the extracellular matrix to which the cells attach. In addition, during angiogenesis endothelial cell migration enables their interaction with other cell types and matrices which may directly affect endothelial cell growth and function. We report the ability to modify the expression of diaphragmed fenestrations and transcapillary channels in capillary endothelial cells by specific extracellular matrices. When the number of membrane openings is measured, growth of the cells on Madin--Darby canine kidney cell matrix results in the greatest number while other matrices or isolated matrix components are only partially active. Production of such a biologically active matrix not only allows an avenue to identify fenestrae-associated proteins but also provides an easy culture method to more closely mimic the in vivo phenotype of endothelial cells.
有窗孔脉管系统中的毛细血管内皮细胞在细胞质的变薄区域含有开口，这些开口通常覆盖有一层或两层隔膜。然而，由于培养时内皮细胞去分化，这种标志性的膜结构常常不表达。更分化表型的表达可由细胞附着的细胞外基质调节。此外，在血管生成过程中，内皮细胞迁移使其能够与其他细胞类型和基质相互作用，这可能直接影响内皮细胞的生长和功能。我们报告了通过特定细胞外基质改变毛细血管内皮细胞中有隔膜窗孔和跨毛细血管通道表达的能力。当测量膜开口数量时，细胞在麦迪逊-达比犬肾细胞基质上生长时产生的膜开口数量最多，而其他基质或分离的基质成分仅具有部分活性。生产这种生物活性基质不仅为鉴定窗孔相关蛋白提供了一条途径，还提供了一种更能紧密模拟内皮细胞体内表型的简便培养方法。
