No abstract

This study proposes a procedure for the isolation and culture of oviduct epithelial cells of several species. In-vitro culture on such a feeder seems to allow full embryonic development and viability. The inner linings of Fallopian tubes from mouse, rabbit, cow and human were trypsinized and the epithelial cells were enriched with Percoll gradient. Isolated cells, obtained in high yield with good viability, were maintained in monolayer culture in B2-Menezo medium supplemented with serum, which also supports early embryonic development in vitro. The plated primary cultures reached confluence within 8 days, producing a monolayer of cohesive polygonal cells. Associated with this large epithelial cell population, ciliated cells as well as polykaryotic cells and few fibroblastic nests were observed. After the first sub-culture, the ciliated cells disappeared and the epithelial cell monolayer grew rapidly to confluence within 3 days and displayed contact inhibition. No epithelial cell growth could be obtained in culture in the absence of serum. The addition of oestrogens had no effect on any of the cultured oviductal epithelial cells. A spontaneous alteration was observed in morphology and growth after several passages, the number of which depends mainly upon the species.

SUMMARY: Bovine epithelial cell monolayers were obtained for culture from fetal oviduct after in situ trypsinization. Isolated ciliated and secretory cells obtained in high yield with good viability were suspended in B2-MENEZO'S medium supplemented with 7.5% fetal bovine serum FBS. The plated primary cultures reached confiuency 2 days after initial seeding, producing a monolayer of cohesive polygonal cells with viability of 85 to 95%. Associated with this large epithelial cell population, ciliated cells as well as a few elongated spindle cells were observed. After the first subculture the ciliated ceils disappeared and the epithelial cells in the monolayer grew more rapidly to confluence than aduh-derived cultures. In addition, frozen-thawed oviduct epithelial cells also maintained a level of 75 to 85% viability during postthaw subculture. The epithelial cells maintained their secretory activity in culture as indicated by electron microscopy and immunocytochemistry. The cell culture monolayers contained keratin, a specific cytoskeletal component of epithelial cells. This culture system may-offer benefits for in ~'itro culture of mammalian embryos.