Modified procedure for isolation of epithelial cells of rat epididymal caput.

Epithelial cells of the rat's epididymal caput were cultivated according to own modification of the Kierszenbaum's method [1981]. The said modification consisted in developing primary cultures of the epithelial cells in the epididymal duct by making use of small tubular segments instead of deisolated cells of the whole epididymal duct wall. Such small segments of the tubules were procured by resorting to mechanical isolation and a 4-grade enzymatic isolation with trypsin and collagenase, whereupon the produced suspension of cells and tubules was filtered through a grid, the meshes of which being 40 X 50 microns in diameter. The cultures were made up exclusively of the tubular segments that had remained on the grid. The utilized technique of isolation gets rid of tubules from the external layer of muscle cells and fibroblasts as well as spermatozoa still prior to the inception of the culture, and provides the possibility to obtain a pure population of epithelial cells. The latter cells have the capacity to migrate from tubular fragments, and to form monolayer cultures. In the conducted cultures the epithelial cells commence secreting PAS-positive substance which was evidenced by means of histochemical and microscope-electron examinations.