Granulosa cells inhibit the resumption of meiosis in bovine oocytes in vitro.
The oocytes of cattle are not as sensitive as those of laboratory animals to purines, cAMP, or follicular extracts. To study the resumption of meiosis, a method is needed that is capable of inhibiting meiosis completely for a minimum of 24 h. This study was designed to evaluate interrelationships in granulosa-oocyte-cumulus complexes using fresh granulosa cells aspirated from small follicles (1-5 mm) in which the cumulus is normally firmly attached. Selected oocyte-cumulus complexes obtained from a slaughterhouse (n = 2,236) were co-incubated with one of the following: various concentrations of fresh granulosa cells in tissue cultures medium (TCM) 199 or bovine follicular fluid (BFF) either without or after one washing and/or freezing; resuspended granulosa cells previously cultured for 7 days; blood cells; or medium alone. Additionally, oocyte-cumulus complexes were embedded in agar cylinders before incubation with or without cells. The rate of maintenance of intact germinal vesicles (GV) in oocytes after 24 h ranged from 40-77% when 5-100 x 10(6) unwashed cells/ml BFF were used, compared to only 16% in oocytes cultured in BFF alone. The pattern was the same when washed cells were used (30-77%, using 5-100 x 10(6) cells/ml BFF), but they were not as effective as unwashed cells. With TCM-199 and the same five concentrations of cells (5, 10, 25, 50, and 100 x 10(6)/ml), a similar inhibition was obtained with greater than or equal to 25 but not with 5 (3%) or 10 (5%) x 10(6)/ml.(ABSTRACT TRUNCATED AT 250 WORDS)
Sirard
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MA
Département de Zootechnie, Université Laval, Ste-Foy, Québec, Canada.
Bilodeau
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S
D000362:Agar; D000818:Animals; D001773:Blood Cells / Q000502:physiology; D002417:Cattle; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D015571:Follicular Fluid / Q000502:physiology; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008540:Meiosis / Q000187:drug effects / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D011687:Purines / Q000494:pharmacology
D011687:Purines; D000362:Agar; D000242:Cyclic AMP