In vitro culture of brushtail possum (Trichosurus vulpecula) epididymal epithelium and induction of epididymal sperm maturation in co-culture

A medium modified from eutherian systems was used to culture epididymal epi thelial cells of the brushtail possum (Trichosurus vulpecula) for more than 2 months. Epididymal tubule fragments from the caput, corpus and cauda epi didymides were used to generate cell monolayers. All three epididymal cell culture systems supported maturational changes in marsupial spermatozoa and enabled immature possum spermatozoa to differentiate from a T-shape to a s treamlined shape, accompanied by the development of progressive motility af ter co-culture with 7-day-old cultured epididymal cell monolayers. This epi didymal cell and sperm co-culture system for marsupial species may facilita te the identification of specific epithelial factors that affect sperm matu ration, particularly in a species in which morphological maturation is read ily visible.