Structural features of cultured epithelial cells from the adult rat epididymis.
Structural features of cultured epithelial cells from the adult rat epididymis.
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DOI：
10.1002/j.1939-4640.1983.tb00761.x
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发表时间：
1983-11
影响因子：
--
通讯作者：
G. Olson;J. Jonas‐Davies;L. H. Hoffman;M. Orgebin‐Crist
中科院分区：
文献类型：
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作者：
G. Olson;J. Jonas‐Davies;L. H. Hoffman;M. Orgebin‐Crist
将从成年大鼠附睾头分离的上皮细胞置于原代培养物中，每天检查外部解剖结构的变化、细胞骨架成分的重组、特征性细胞质特征的维持以及对培养基的反应，以最大限度地减少非上皮细胞增殖。在培养的第一个24小时后，细胞开始显著附着于基质。贴壁后，细胞逐渐变平，并紧密贴附在基底上。这是伴随着细胞表面上的微绒毛的重新分布和细胞内的细胞骨架元素的重组。扁平化后，培养的细胞显示出广泛的阵列的10 nm的细丝，这是与桥粒连接相邻的细胞。免疫荧光研究表明，这些是含有角蛋白的中间丝和2-D凝胶电泳的完整细胞和细胞骨架显示，一个家庭的“角蛋白样”多肽的细胞的主要成分。原代培养中的上皮细胞附着、形态和维持不受D-缬氨酸、阿糖胞苷或两者的影响;然而，这些药物单独或联合使用，显著减少了掺入3 H-胸苷的细胞数量。这些数据表明，分离的上皮细胞保留了一些分化的结构特征，这些特征是完整细胞的特征，并且可以在成纤维细胞增殖被抑制的条件下维持上皮细胞的富集培养物。
Epithelial cells isolated from the caput epididymidis of adult rats were placed in primary culture and examined daily for ten days for changes in external anatomy, reorganization of cytoskeletal components, maintenance of characteristic cytoplasmic features, and response to media formulated to minimize nonepithelial cell proliferation. Significant cell attachment to the substrate began after the first 24 hours of culture. After attachment, the cells underwent a progressive flattening and became closely applied to the substrate. This was accompanied by a redistribution of microvilli on the cell surface and a reorganization of cytoskeletal elements within the cell. After flattening, the cultured cells displayed an extensive array of 10-nm filaments which were associated with the desmosomes attaching adjacent cells. Immunofluorescence studies demonstrated that these were keratin-containing intermediate filaments and 2-D gel electrophoresis of intact cells and cell cytoskeletons revealed that a family of "keratin-like" polypeptides were major components of the cells. Epithelial cell attachment, morphology, and maintenance in the primary culture were unaffected by D-valine, cytosine arabinoside, or both; however, these agents, either individually or in combination, reduced significantly the number of cells incorporating 3H-thymidine. These data show that isolated epithelial cells retain some differentiated structural features that characterize the intact cell and that enriched cultures of epithelial cells can be maintained under conditions where fibroblast proliferation is inhibited.