Yue B Y, Sugar J, Gilboy J E, Elvart J L
Department of Ophthalmology, Lions of Illinois Eye Research Institute, University of Illinois, Chicago 60612.
Invest Ophthalmol Vis Sci. 1989 Feb;30(2):248-53.
We investigated the effects of various culture conditions on the growth of normal human corneal endothelial cells in culture. Falcon Primaria tissue culture plastic was found to provide a more suitable surface for endothelial cell growth than the conventional Corning tissue culture plastic. Also, media containing 10% fetal bovine serum and 5% calf serum (complete media) facilitated the growth of human cells better than those containing Nu-serum. Supplementation with epidermal or fibroblast growth factor (10 and 100 ng/ml) to the complete media had no effect on human endothelial cell growth. Chondroitin sulfate at low concentrations (100 micrograms/ml to 1 mg/ml) also showed little effect. At high concentrations (13.5 and 25 mg/ml), however, chondroitin sulfate significantly promoted human corneal endothelial cell growth during a 1- to 2-week incubation period. From the 37 cultures initiated, outgrowth from explants appeared within 3 to 7 days. Cells were polygonal in shape and, at confluency, formed a continuous monolayer. We attained a success rate of 87% (7/8) growing primary cultures from donors under 20 years of age and a 59% (17/29) success rate from older donors.
我们研究了各种培养条件对培养的正常人角膜内皮细胞生长的影响。发现Falcon Primaria组织培养塑料比传统的康宁组织培养塑料为内皮细胞生长提供了更合适的表面。此外，含有10%胎牛血清和5%小牛血清的培养基（完全培养基）比含有新生牛血清的培养基更有利于人类细胞的生长。向完全培养基中添加表皮生长因子或成纤维细胞生长因子（10和100 ng/ml）对人内皮细胞生长没有影响。低浓度（100微克/毫升至1毫克/毫升）的硫酸软骨素也显示出几乎没有影响。然而，在高浓度（13.5和25毫克/毫升）下，硫酸软骨素在1至2周的孵育期内显著促进了人角膜内皮细胞的生长。在开始的37个培养物中，外植体在3至7天内出现生长。细胞呈多边形，汇合时形成连续的单层。我们从20岁以下供体的原代培养物中获得了87%（7/8）的成功率，从老年供体中获得了59%（17/29）的成功率。
