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在器官培养系统中,对生长在明胶海绵基质上的人视网膜色素上皮细胞、视网膜神经胶质细胞和成纤维细胞进行的形态学和免疫组织化学研究。结构蛋白和非结构蛋白的比较及其在细胞类型鉴定中的应用。

A morphological and immunohistochemical study of human retinal pigment epithelial cells, retinal glia, and fibroblasts grown on Gelfoam matrix in an organ culture system. A comparison of structural and nonstructural proteins and their application to cell type identification.

作者信息

Vinores S A, Herman M M, Hackett S F, Campochiaro P A

机构信息

Department of Ophthalmology, University of Virginia School of Medicine, Charlottesville 22908.

出版信息

Graefes Arch Clin Exp Ophthalmol. 1993 May;231(5):279-88. doi: 10.1007/BF00919106.

Abstract

Retinal pigment epithelial (RPE) cells, retinal glia, and fibroblasts undergo marked phenotypical change when outside their usual microenvironment, as occurs in epiretinal membrane formation. To explore their phenotypic potential without the influence of other cell types, each was cultured on Gelfoam matrix and assessed immunohistochemically and ultrastructurally. All cell types demonstrated vimentin and a universal beta-tubulin epitope, TU27. RPE and retinal glial cells were positive for cytokeratin, Leu 7, and neuron-specific (gamma gamma) enolase, as were glia and fibroblasts for S-100 protein and RPE cells and fibroblasts for glutamine synthetase. RPE cells alone showed positivity for class III beta-tubulin and retinal S-antigen (monolayer cultures only); occasional retinal glia, which immunohistochemical findings suggest are Müller cell derived, demonstrated GFA protein. Therefore, class III beta-tubulin may be useful in distinguishing RPE cells from retinal glia and fibroblasts, and Leu-7 may help to identify RPE cells and fibroblasts; these cell types are difficult to distinguish in clinical material using more traditional morphological criteria.

摘要

视网膜色素上皮(RPE)细胞、视网膜神经胶质细胞和成纤维细胞在脱离其正常微环境时会发生显著的表型变化,视网膜前膜形成时就会出现这种情况。为了在不受其他细胞类型影响的情况下探索它们的表型潜能,将每种细胞培养在明胶海绵基质上,并进行免疫组织化学和超微结构评估。所有细胞类型均显示波形蛋白和一种通用的β-微管蛋白表位TU27。RPE细胞和视网膜神经胶质细胞的细胞角蛋白、Leu 7和神经元特异性(γγ)烯醇化酶呈阳性,神经胶质细胞和成纤维细胞的S-100蛋白呈阳性,RPE细胞和成纤维细胞的谷氨酰胺合成酶呈阳性。仅RPE细胞的III类β-微管蛋白和视网膜S抗原呈阳性(仅单层培养);偶尔有视网膜神经胶质细胞,免疫组织化学结果表明其来源于Müller细胞,显示胶质纤维酸性蛋白(GFA蛋白)阳性。因此,III类β-微管蛋白可能有助于区分RPE细胞与视网膜神经胶质细胞和成纤维细胞,Leu-7可能有助于识别RPE细胞和成纤维细胞;使用更传统的形态学标准在临床材料中很难区分这些细胞类型。

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