Effect ofd-valine and cytosine arabinoside on [3H]thymidine incorporation in rat and rabbit epididymal epithelial cell cultures

Summary Epithelial cell enriched primary cultures were established from the rat and the rabbit epididymis. Epithelial cell aggregates, obtained after pronase digestion of minced epididymis, attached to the culture dish and after 72 h in vitro spread out to form discrete patches of cells. These cells have an epithelioid morphology and form a monolayer of closely apposed polygonal cells where DNA synthesis, as judged by [³H]thymidine uptake, is very low. Inl-valine medium the nonepithelial cell contamination was no more than 10% in rat and rabbit epididymal primary cultures. The labeling index of rat epididymal cells cultured ind-valine medium was significantly lower than that of cells cultured inl-valine medium. In contrast, the labeling index of rabbit epididymal cells cultured ind-valine medium was significantly higher than that of cells cultured inl-valine medium. Cytosine arabinoside decreased the number of labeled cells in bothl-valine andd-valine cultures. From these results, it appears thatd-valine is a selective agent for rat epididymal epithelial cells, but not for rabbit epithelial cells, and that cytosine arabinoside is a simple and effective means to control the proliferation of fibroblast-like cells in both rat and rabbit epididymal cell cultures. This research was sponsored by grants from the National Institute of Child Health and Human Development, Bethesda, MD (HD-03820, HD-11816, HD-05797), and the Mellon Foundation.