McIntosh L C, Muckersie L, Forrester J V
Department of Ophthalmology, University of Aberdeen.
Tissue Cell. 1988;20(2):193-209. doi: 10.1016/0040-8166(88)90041-9.
Recent studies have shown that the extracellular matrix modifies the behaviour of endothelial cells. We have studied the effects of extracellular matrix components on retinal capillary endothelial cell migration and proliferation. Bovine retinal capillary endothelial cells were selectively cultured from collagenase-digested microvessel fragments. In a filter system for the assessment of migration, endothelial cells responded to substrate-bound fibronectin but not to soluble fibronectin. Cell migration on collagen- or gelatin-coated filters was minimal, and these cells failed to adopt a spread morphology, remaining instead as round cells. Cell replication was quantified using a protein dye binding assay for adherent cells in 96 well plates. Serum was essential for growth irrespective of the substrate. Cells harvested from microvessel cultures proliferated more rapidly on collagen- and gelatin-coated plastic than on fibronectin and were unaffected by additions to the medium such as endothelial cell conditioned medium, whereas cells proliferating directly from the microvessels grew at a faster rate on fibronectin and also responded to conditioned medium supplement. When cultured on collagen gels, initial microvessel cells and harvested cells required surface fibronectin in order to adopt a cobblestone morphology. These results show that fibronectin is a requirement for bovine retinal capillary endothelial cell migration, but proliferation of these cells can be supported, with slight differences, by both fibronectin and collagen provided serum growth factors are present. These findings are relevant to the early phase of angiogenesis in which migration and proliferation of endothelial cells occurs.
最近的研究表明，细胞外基质可改变内皮细胞的行为。我们研究了细胞外基质成分对视网膜毛细血管内皮细胞迁移和增殖的影响。从胶原酶消化的微血管片段中选择性培养牛视网膜毛细血管内皮细胞。在用于评估迁移的滤膜系统中，内皮细胞对结合于底物的纤连蛋白有反应，但对可溶性纤连蛋白无反应。在胶原或明胶包被的滤膜上细胞迁移极少，且这些细胞无法形成铺展形态，而是保持圆形细胞状态。使用蛋白质染料结合测定法对96孔板中的贴壁细胞进行细胞复制定量。无论底物如何，血清对生长都是必不可少的。从微血管培养物中收获的细胞在胶原和明胶包被的塑料上比在纤连蛋白上增殖更快，并且不受培养基添加物如内皮细胞条件培养基的影响，而直接从微血管增殖的细胞在纤连蛋白上生长更快，并且也对条件培养基补充物有反应。当在胶原凝胶上培养时，最初的微血管细胞和收获的细胞需要表面纤连蛋白才能形成鹅卵石样形态。这些结果表明，纤连蛋白是牛视网膜毛细血管内皮细胞迁移所必需的，但只要存在血清生长因子，纤连蛋白和胶原都能在一定程度上支持这些细胞的增殖。这些发现与内皮细胞发生迁移和增殖的血管生成早期阶段相关。
