title: In vitro culture of epithelial cells from the caput, corpus, and cauda epididymis of Sus domesticus.
abstract: This work describes a protocol to culture epididymal epithelial cells from the caput, corpus, and cauda regions of Sus domesticus. Epididymal epithelial fragments were obtained by dissection and enzymatic digestion with collagenase. About 30 epididymal fragments from each epididymal region were cultured in 24-well culture plates with supplemented RPMI-1640 medium at 37 degrees C, 5% CO2 in air, and 100% humidity. A confluent monolayer of polygonal and tightly packed epithelioid cells from the three epididymal regions was obtained after 12-16 days in culture and maintained in vitro for more than 60 days. The proportion of epididymal epithelial cells in these cultures was assessed by immunofluorescent staining for cytokeratins. Throughout the 2 months of culture, about 80% of the cells were cytokeratin-positive. Electron microscopy observations indicated that cultured cells from caput, corpus, and cauda epididymal regions were tightly adhered to each other by junctional complexes and that stereocilia were present in their apical membranes. Moreover, the presence of an extensive rough endoplasmic reticulum, Golgi apparatus and numerous vesicles in the cytoplasm suggested that cultured cells maintained secretory and absorptive activities. These results show that the epididymal epithelial cells in culture from S. domesticus retain some fundamental features that characterize the epididymal epithelium in the intact organ. This system might be a valuable tool for studying the mechanism of sperm maturation in vitro, including epididymal cell secretions and the analysis of regional differences.
authors.affiliation: Biotechnology of Porcine Reproduction, Department of Biology, Faculty of Sciences, University of Girona, Campus Montilivi sn, 17071, Spain. judit.bassols@udg.es
authors.forename: Judit
authors.initials: J
authors.lastname: Bassols
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kádár
authors.forename: M Dolors
authors.initials: MD
authors.lastname: Briz
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Pinart
authors.forename: Sílvia
authors.initials: S
authors.lastname: Sancho
authors.forename: Núria
authors.initials: N
authors.lastname: Garcia-Gil
authors.forename: Elena
authors.initials: E
authors.lastname: Badia
authors.forename: Anna
authors.initials: A
authors.lastname: Pruneda
authors.forename: Eva
authors.initials: E
authors.lastname: Bussalleu
authors.forename: Marc
authors.initials: M
authors.lastname: Yeste
authors.forename: Sergi
authors.initials: S
authors.lastname: Bonet
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002452:Cell Count; D018929:Cell Culture Techniques / Q000662:veterinary*; D002455:Cell Division; D004822:Epididymis / Q000166:cytology*; D004847:Epithelial Cells / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007633:Keratins / Q000032:analysis; D008297:Male; D008854:Microscopy, Electron; D034421:Sus scrofa
chemical_list: D007633:Keratins