Growth and characterization of polarized monolayers of epididymal epithelial cells and Sertoli cells in dual environment culture chambers.
Growth and characterization of polarized monolayers of epididymal epithelial cells and Sertoli cells in dual environment culture chambers.
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DOI：
10.1002/j.1939-4640.1986.tb00871.x
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发表时间：
1986-01
影响因子：
--
通讯作者：
S. W. Byers;M. A. Hadley;D. Djakiew;Martin Dym
中科院分区：
文献类型：
--
作者：
S. W. Byers;M. A. Hadley;D. Djakiew;Martin Dym
将从成熟大鼠分离的附睾上皮细胞和从10日龄大鼠分离的支持细胞在无血清限定培养基中培养于细胞外基质浸渍的过滤器上，该过滤器保持在双环境培养室中。附睾上皮细胞只有在高密度（大于1 × 10（6）个细胞/cm 2）下接种时才出现极化现象。这些细胞的汇合单层形成了菊粉的渗透屏障。支持细胞是柱状和高度极化的细胞外基质浸渍过滤器上生长时，立方形的过滤器单独生长时，和鳞状塑料上生长时。这些细胞的汇合极化单层排除了电子致密示踪硝酸镧的基底紧密连接的方式。因此，极化的附睾上皮细胞和支持细胞的单层可以通过在细胞外基质浸渍的可渗透支持物上以高密度生长细胞来获得。通过在特殊构造的培养室中保持单层，细胞可以形成渗透性屏障，并且能够实现顶端室与基底室的分离，这对于它们在体内的功能非常重要。
Epididymal epithelial cells isolated from mature rats and Sertoli cells isolated from 10-day-old rats were cultured in serum-free defined media on extracellular matrix impregnated filters maintained in dual environment culture chambers. Epididymal epithelial cells had a polarized appearance only when plated at high density (greater than 1 X 10(6) cells/cm2). Confluent monolayers of these cells formed a permeability barrier to inulin. Sertoli cells were columnar and highly polarized when grown on extracellular matrix-impregnated filters, cuboidal when grown on filters alone, and squamous when grown on plastic. Confluent polarized monolayers of these cells excluded the electron-dense tracer lanthanum nitrate by way of basal-tight junctions. Therefore, polarized monolayers of epididymal epithelial cells and Sertoli cells can be obtained by growing the cells at high density on extracellular matrix-impregnated permeable supports. By maintaining the monolayers in specially constructed culture chambers, the cells can develop a permeability barrier, and are able to achieve the separation of apical from basal compartments so important for their function in vivo.