Isolation, culture and characteristics of epididymal epithelial cells from adult cats.
A tissue-culture system in which cells retain defined ultrastructural and functional characteristics was established to provide a basis for functional investigations of the epididymal duct in the cat. A widely used culture protocol for rat epididymal epithelium was used as a starting point and subsequently modified. The cellular population of the cat's epididymal epithelium was isolated by successive collagenase and trypsin digestion. A high yield of isolated cells obtained with good viability, were cultured in DMEM/F12 medium supplemented with foetal bovine serum, in absence or in presence of additional dihydrotestosterone (1 nM). The plated primary cultures reached confluence within 5-8 days, producing a monolayer of cohesive cells. Samples taken after 6 days in culture were processed for transmission and scanning electron microscopies. Immunocytochemical staining was used to estimate the purity of the epithelial cell population in the monolayers. The cell cultures displayed several functional traits of in vivo epithelia, including [35S] hypotaurine and [35S] taurine production. These results demonstrate that primary cultures of epididymal epithelial cells isolated from sexually mature cats maintain several differentiated characteristics of the intact organ and therefore provide a valuable system for the study of epididymal epithelial cell functions, metabolic activities and their regulation in cats.
Buff
Samuel
S
Unité de Biologie de la Reproduction, Ecole Nationale Vétérinaire de Lyon, 1, Avenue Bourgelat, F-69280 Marcy l'Etoile, France. s.buff@vet-lyon.fr
Lambert
Véronique
V
Marchal
Thierry
T
Guérin
Pierre
P
D000818:Animals; D002415:Cats; D002455:Cell Division; D002469:Cell Separation; D002478:Cells, Cultured; D013196:Dihydrotestosterone / Q000494:pharmacology; D004822:Epididymis / Q000166:cytology*; D004847:Epithelial Cells / Q000648:ultrastructure*; D007150:Immunohistochemistry; D008297:Male; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning
D013196:Dihydrotestosterone