Biology of Reproduction, Vol 15, 266-279, Copyright © 1976 by Society for the Study of Reproduction

Isolation of Principal and Basal Cells from the Epithelium of the Hamster Caput Epididymidis by Unit Gravity Sedimentation

¹ Department of Dairy and Animal Science, The Pennsylvania State University, University Park, PA 16802
² Department of Biology, The Pennsylvania State University, University Park, PA 16802

A technique was developed to enzymatically disaggregate cells comprising the caput epididymidis. The resulting mixture of dispersed cells was separated by sedimentation in a shallow bovine serum albumin gradient at unit gravity into populations of spermatozoa, erythrocytes and several nucleated cell types. Separation by this method primarily was on the basis of cell size. Light and electron microscopy were used to identify the separated somatic cell types and to determine the homogeneity of each population. The purest fractions of the six populations (from smallest to largest) contained an average of 84 percent erythrocytes, 76 percent basal cells, 82 percent fibroblasts and intraepithelial lymphocytes, 68 percent small principal cells, 34 percent smooth muscle cells or 58 percent large principal cells. Based on trypan blue exclusion, ATP content and fine structure in electron micrographs we concluded that cell viability following sedimentation was excellent. Isolated epithelial cells made available by this technique should enable critical analyses of epididymal function.

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ACKNOWLEDGMENTS This research was supported by NIH grant HD-07244 and authorized for publication as Paper No. 4999 in the journal series of the Pennsylvania Agricultural Experiment Station. We are indebted to Mr. H. M. Schuler and Drs. R. H. Hammerstedt, W. C. Hymer and D. W. Hamilton for comments and suggestions during the course of this study. Mrs. J. Eastman provided expert technical assistance.