Isolation of principal and basal cells from the epithelium of the hamster caput epididymidis by unit gravity sedimentation

A technique for the isolation of principal and basal cells from the epithelium of the hamster caput epididymides by unit gravity sedimentati on is described. The technique enzymatically disaggregates cells comprising the caput epididymides, and the resulting mixture of disperse d cells is separated by sedimentation in a shallow bovine serum albumin gradient at unit gravity into populations of spermatozoa, erythrocytes and several nucleated types. The separated somatic cell types and the homogeneity of each population were identified by light and electron microscopy. The purest fractions of the 6 populations, from smallest to largest, contained an average of 84% erythrocytes, 76% basal cells, 82% fibroblasts and intraepithelial lymphocytes, 68% small principal cells and 34% smooth muscle cells or 58% large principal cells. Cell viability following sedimentation was excellent, as concluded from elect ron micrographs revealing adenosine triphosphate content and fine struct ure. This technique should enable critical analyses of epididymal function in isolated epithelial cells.