DEVELOPMENT OF CELL-SURFACE POLARITY IN THE EPITHELIAL MADIN-DARBY CANINE KIDNEY (MDCK) CELL-LINE
DEVELOPMENT OF CELL-SURFACE POLARITY IN THE EPITHELIAL MADIN-DARBY CANINE KIDNEY (MDCK) CELL-LINE
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DOI：
10.1002/j.1460-2075.1984.tb02194.x
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发表时间：
1984-01-01
期刊：
影响因子：
11.4
通讯作者：
SIMONS, K
中科院分区：
文献类型：
--
作者：
BALCAROVASTANDER, J;PFEIFFER, SE;SIMONS, K
通过检查两种基底侧标记物（抗58-kd蛋白的单克隆抗体和[35 S]蛋氨酸摄取），在上皮Madin达比犬肾（MDCK）细胞系中研究了表面极性的发展。将细胞接种在盖玻片或硝酸纤维素滤膜上后，跟踪这些标记物的表面分布。在亚融合单层中，许多细胞的顶端表面被抗58-kd抗体染色。在盖玻片上生长的细胞达到汇合后，首先发生顶端表面的清除。同样，在滤器上生长的细胞中，基底外侧58-kd蛋白从顶面消失，同时在细胞单层上产生可测量的电阻。从过滤器生长细胞的两侧测量[35 S]蛋氨酸的摄取，并在接种后早期开始增加，在完全极化的单层中达到大于98%基底外侧的值。这些结果强调了MDCK细胞表面极性的发展是一个渐进的过程，并且完整的表面极化似乎需要广泛的细胞-细胞接触。
The development of surface polarity has been studied in the epithelial Madin‐Darby canine kidney (MDCK) cell line by examining two basolateral markers: a monoclonal antibody against a 58‐kd protein and [35S]methionine uptake. The surface distribution of these markers was followed after plating the cells on coverslips or nitrocellulose filters. In subconfluent monolayers the apical surface of many cells was stained with the anti‐58‐kd antibody. Clearing of the apical surface occurred first after confluency had been reached in cells grown on coverslips. Similarly, in cells grown on filters the basolateral 58‐kd protein disappeared from the apical surface concomitantly with the development of a measurable electrical resistance over the cell monolayer. The uptake of [35S]methionine was measured from both sides of filter‐grown cells and began to polarize early after seeding, reaching a value of greater than 98% basolateral in the fully polarized monolayer. These results emphasize that the development of surface polarity in MDCK cells is a gradual process, and that extensive cell‐cell contacts seem to be required for complete surface polarization.