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Bovine granulosa and oviductal cell secretions support sperm motility
Theriogenology
39:234,
1993
BOVINE GRANULOSA AND OVIDUCTAL CELL SECRETIONS SUPPORT SPERM MGHLlTY A. Ijaz* and M. A. Sirard Dkpartement de zootechnie, Laboratoire d’Gntogenie Reproduction du Centre de Rechetche du CHUL, Universite Laval, Quebec, Canada GlK 7P4. The present study was designed to examine the effect of bovine granulosa and oviductal cell secretions on sperm motility in vitro. Granulosa cells (GC;from l-5 mm follicles) and oviductal cells (0C;from proestrus and estrus oviducts) were collected from reproductive tracts obtained from a slaughter house, and were cultured for 3 days in TCM-199 medium supplemented with 10% fetal calf serum in 50 ml culture bottles. For GC, the supematant was discarded and the monolayer was covered with TL sperm medium containing 6 m&nl bovine serum albumin (TL-SP). The OC were recovered by centrifugation, transferred to culture bottles containing TL-SP and were cultured for further 2 days. On Day 5 , GC conditioned and GC conditioned TL-SP were recovered, frozen (-20 C), and dialysed (dialysis tubing cut off 6000 to 8000 MW). The dialysates (D) and retentates (R) were lyophilized or used directly for motility experiments. Bovine follicular fluid (BFF$rom l5 mm follicles) was also dialysed and the D and R were lyophilized. In the first experiment, frozen thawed sperm (Given by CIAQJ were washed twice in TL-SP and incubated in TL-SP, GC conditioned TL-SP or GC conditioned TL-SP for 30 hr. The data were analysed by ANOVA. Motility decreased @
234
Copyright 0 1993 Buttervvorth-Heinemann
39:234,
1993
BOVINE GRANULOSA AND OVIDUCTAL CELL SECRETIONS SUPPORT SPERM MGHLlTY A. Ijaz* and M. A. Sirard Dkpartement de zootechnie, Laboratoire d’Gntogenie Reproduction du Centre de Rechetche du CHUL, Universite Laval, Quebec, Canada GlK 7P4. The present study was designed to examine the effect of bovine granulosa and oviductal cell secretions on sperm motility in vitro. Granulosa cells (GC;from l-5 mm follicles) and oviductal cells (0C;from proestrus and estrus oviducts) were collected from reproductive tracts obtained from a slaughter house, and were cultured for 3 days in TCM-199 medium supplemented with 10% fetal calf serum in 50 ml culture bottles. For GC, the supematant was discarded and the monolayer was covered with TL sperm medium containing 6 m&nl bovine serum albumin (TL-SP). The OC were recovered by centrifugation, transferred to culture bottles containing TL-SP and were cultured for further 2 days. On Day 5 , GC conditioned and GC conditioned TL-SP were recovered, frozen (-20 C), and dialysed (dialysis tubing cut off 6000 to 8000 MW). The dialysates (D) and retentates (R) were lyophilized or used directly for motility experiments. Bovine follicular fluid (BFF$rom l5 mm follicles) was also dialysed and the D and R were lyophilized. In the first experiment, frozen thawed sperm (Given by CIAQJ were washed twice in TL-SP and incubated in TL-SP, GC conditioned TL-SP or GC conditioned TL-SP for 30 hr. The data were analysed by ANOVA. Motility decreased @
234
Copyright 0 1993 Buttervvorth-Heinemann