Capetandes A, Gerritsen M E
Department of Physiology, New York Medical College, Valhalla 10595.
Invest Ophthalmol Vis Sci. 1990 Sep;31(9):1738-44.
Different matrix components, in combination with various media and serum supplements, were evaluated for their ability to promote selectively the growth of bovine retinal endothelial cells in primary culture. The optimal setting for the selective growth of retinal endothelial cells was a fibronectin/hyaluronic acid matrix, Dulbecco's modified Eagle's medium (DMEM) supplemented with 20% pooled human serum and 100 micrograms/ml heparin. These conditions consistently yielded virtually homogeneous cultures of endothelial cells, assessed using specific endothelial markers. Thus obtained, the retinal endothelial cells could be subcultured and maintained in phenotypically stable long-term serial cultivation. Homogeneous cultures of retinal pericytes were obtained when microvessel isolates were seeded to uncoated or gelatin-coated culture dishes and grown in DMEM supplemented with 20% fetal bovine serum. The retinal pericytes could also be subcultured and cultivated for numerous populations doublings. Additionally, observations from this study suggest that two populations of pericytes may be obtained in culture and distinguished on the basis of their relative size and antigenic properties.
评估了不同的基质成分与各种培养基和血清补充剂组合后，促进原代培养的牛视网膜内皮细胞选择性生长的能力。视网膜内皮细胞选择性生长的最佳条件是纤连蛋白/透明质酸基质、补充有20%混合人血清和100微克/毫升肝素的杜尔贝科改良伊格尔培养基（DMEM）。使用特异性内皮标记物评估，这些条件始终能产生几乎均一的内皮细胞培养物。如此获得的视网膜内皮细胞可以传代培养，并在表型稳定的长期连续培养中维持。当将微血管分离物接种到未包被或明胶包被的培养皿中，并在补充有20%胎牛血清的DMEM中培养时，可获得视网膜周细胞的均一培养物。视网膜周细胞也可以传代培养并进行多次群体倍增培养。此外，本研究的观察结果表明，在培养中可能获得两种周细胞群体，并可根据它们的相对大小和抗原特性加以区分。
