Isolation and Cell Culture of the Epithelial Cells of Cauda Epididymidis of the Bull1

A simple and rapid method has been described to isolate the epithelial cells of cauda epididymidis of adult bull. Perfusion of the lumen of the cauda epididymidis with 1 mg/ml collagenase in calcium- and magnesium-free Hank's balanced salt solution and incubation of the tissue at 37 degrees C for 90 min releases the principal and basal cells into the lumen. Several individual epithelial cells and cell aggregates without the contamination of stromal or smooth muscle cells can be flushed out at the end of incubation. The isolated epithelial cells, suspended in Dulbecco's medium with 10% horse serum, attach to plastic dishes within 3 h after seeding the cells and proliferate to form a monolayer in approximately 8-12 days. The electron microscopic study and immunostaining of the cultured epithelial cells indicate that the cultured cells are principal cells. The basal cells of the intact cauda epididymidis of bull show within their cytoplasm the presence of varying amounts of "lipid-like" material often closely associated with whorls of membrane.