Sperm-Oviduct Interaction: MATERIALS AND METHODS(1)

OEC Preparation

Oviducts were obtained at a local slaughterhouse from prepubertal gilts of ~120 days of age. The ovaries did not show any signs of cyclicity such as follicular growth, ovulation, or the presence of corpora lutea, and they were covered with small follicles (2-3-mm diameter). Oviducts were transported to the laboratory in PBS at room temperature (22°C). Upon arrival, the oviducts were cleaned and washed using PBS. They were trimmed from the ovaries, transferred to the washing medium containing Hanks’ Balanced Salt Solution (Life Technologies, Paisley, UK) supplemented with 100 U/ml penicillin, 100 ^g/ml streptomycin, and 0.25 ^g/ml amphotericin B (Life Technologies), and then gently rinsed.

Each oviduct was divided into three sections; one between the fimbria and the middle of the ovi-ductal tube, containing the thicker part of the oviduct tube, was designated as ampulla. A section containing 1-2 cm of the caudal part of the uterus horn, uterotubal junction, and up to nearly the middle of the oviductal tube containing the thinner part of the oviduct tube was designated as isthmus. Finally, a section around the junction of the thin and thick part of the oviductal tube, approximately 1-2 cm long, was cut and discarded, to assure differentiation of isthmic and ampullar parts of the oviduct. The isthmic and ampullar sections of oviduct were opened longitudinally. The epithelial cells were scraped using the blunt side of a scalpel blade.

This entry was posted in Sperm and tagged Epithelial Cells, Oviduct, Sperm, Spermatozoa.