Cytoskeleton, adhesion, and extracellular matrix of fetal human retinal pigmented epithelial cells in culture.
Cytoskeleton, adhesion, and extracellular matrix of fetal human retinal pigmented epithelial cells in culture.
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培养物中胎儿视网膜色素上皮细胞的细胞骨架、粘附和细胞外基质。
DOI：
10.1159/000266768
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发表时间：
1989
影响因子：
2.1
通讯作者：
V. Kalnins
中科院分区：
文献类型：
--
作者：
K. Turksen;M. Opas;V. Kalnins
取6~9周龄人胎眼视网膜色素上皮(RPE)细胞，建立RPE细胞培养体系。培养4-5天后，免疫荧光和荧光显微镜观察集落不同部位的RPE细胞骨架蛋白、微管蛋白、肌球蛋白、F-肌动蛋白、波形蛋白和细胞角蛋白的分布以及细胞外基质蛋白层粘连蛋白和IV型胶原的分布。细胞外基质成分层粘连蛋白和IV型胶原大量存在于集落中心的RPE细胞下，向集落边缘逐渐减少。小管蛋白抗血清标记的微管分布于RPE细胞胞浆内。F-肌动蛋白和肌球蛋白存在于集落边缘RPE细胞的应力纤维中和集落中心RPE细胞顶端周围致密的环状结构中。干涉反射显微镜对细胞-基质黏附模式的检测表明，焦点接触仅存在于位于集落边缘的RPE细胞中，它们位于应力纤维的末端。然而，以前在鸡RPE细胞集落中观察到的大灶性粘连在人RPE细胞中没有看到。整个集落中的人RPE细胞均含有含细胞角蛋白的中间纤维和含波形蛋白的中间纤维，而在鸡RPE细胞中仅观察到后者。这些结果表明，在细胞骨架的一般组织结构和细胞外基质成分、层粘连蛋白和IV型胶原的分布上，人RPE细胞的集落与鸡RPE细胞的集落相似，但不完全相同。
Small sheets of human retinal pigmented epithelial (RPE) cells isolated from 6- to 9-week-old fetal human eyes were used to establish RPE cells in culture. After 4-5 days in culture the RPE cells in various parts of a colony were examined by immunofluorescence and fluorescence microscopy to determine the distribution of the cytoskeletal proteins tubulin, myosin, F-actin, vimentin, and cytokeratin and the extracellular matrix proteins laminin and type IV collagen. The extracellular matrix components laminin and type IV collagen, which are abundant underneath the RPE cells in the centre of the colony, progressively decrease in amount towards the edge of the colony. Microtubules labeled by antisera to tubulin were present in the cytoplasm of RPE cells throughout the colony. F-actin and myosin were found in stress fibres in the RPE cells at the edge of the colony and in compact circumferential rings circumscribing the apical ends in RPE cells in the centre of the colony. Examination of cell-substratum adhesion patterns by interference reflection microscopy showed that focal contacts were present only in RPE cells at the edge of the colony where they were located at the ends of the stress fibres. The large focal adhesions previously observed in colonies of chick RPE cells, however, were not seen in human RPE cells. Human RPE cells throughout the colony had cytokeratin-containing intermediate filaments as well as vimentin-containing intermediate filaments, whereas only the latter were observed in chick RPE cells. These results indicate that in the general organization of the cytoskeleton and in the distribution of the extracellular matrix components, laminin and type IV collagen, colonies of human RPE cells are similar but not identical to colonies of chick RPE cells.