title: Growth and characterization of polarized monolayers of epididymal epithelial cells and Sertoli cells in dual environment culture chambers.
abstract: 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.
mesh_terms: D000818:Animals; D002452:Cell Count; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003470:Culture Media; D004822:Epididymis / Q000502:physiology* / Q000648:ultrastructure; D004847:Epithelial Cells; D005109:Extracellular Matrix; D007365:Intercellular Junctions / Q000502:physiology; D007444:Inulin; D007811:Lanthanum; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D012708:Sertoli Cells / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D003470:Culture Media; C016534:lanthanide nitrate; D007811:Lanthanum; D007444:Inulin