Marceau F, Boisjoly H M, Wagner E, Lille S, Roy R
Unité de recherche Inflammation et Immunologie-Rhumatologie, Centre Hospitalier de l'Université Laval, Ste-Foy, Québec, Canada.
Exp Eye Res. 1990 Dec;51(6):645-50. doi: 10.1016/0014-4835(90)90048-y.
Human limbal explants obtained from 44 eyebank donors were cultured in medium 199 supplemented with 20% fetal bovine serum, vascular endothelial cell growth supplement and heparin. Cells grew abundantly out of the explants. They initially formed a 'cobblestone' patterned monolayer but later exhibited an elongated morphology with growth in parallel bundles. These cells could be passaged at least nine times and were identified throughout the consecutive passages as microvascular endothelial cells by the expression of factor VIII-related antigen, laminin and of the H determinant of ABO blood groups. As expected from vascular endothelial cells, flow cytometric analysis demonstrated a strong expression of class I histocompatibility antigens and a weaker expression of class II antigens. Class II antigen expression was enhanced by culturing the cells in the presence of immune interferon. These cells produced immunoreactive interleukin-1, mainly of the alpha type, under endotoxin stimulation. Limbal microvascular cells could be useful to study corneal angiogenesis. Furthermore, long-term culture of limbal cadaveric tissue can potentially be used to characterize donor-specific immunologic responses in corneal graft recipients.
从44位眼库供体获取的人角膜缘外植体在添加了20%胎牛血清、血管内皮细胞生长因子和肝素的199培养基中培养。细胞从外植体大量生长出来。它们最初形成“鹅卵石”样排列的单层，但后来呈现出细长形态并平行成束生长。这些细胞至少可传代9次，在连续传代过程中通过VIII因子相关抗原、层粘连蛋白和ABO血型H抗原的表达被鉴定为微血管内皮细胞。正如血管内皮细胞所预期的那样，流式细胞术分析显示I类组织相容性抗原强烈表达，II类抗原表达较弱。在免疫干扰素存在的情况下培养细胞可增强II类抗原表达。在内毒素刺激下，这些细胞产生免疫反应性白细胞介素-1，主要是α型。角膜缘微血管细胞可能有助于研究角膜血管生成。此外，角膜缘尸体组织的长期培养有可能用于表征角膜移植受者的供体特异性免疫反应。
