Fertilization and early cleavage in vitro of ageing bovine oocytes after maturation in culture

Bovine follicular oocytes cultured for 20 to 48 hours were inseminated with frozen thawed spermatozoa. Significantly higher penetration rates (94 to 100% vs 86 to 94%: P < 0.05) and proportions of polyspermy (35 to 78% vs 22 to 45%: P < 0.01) were obtained for oocytes cultured continuously for 24 hours with spermatozoa than for those separated from spermatozoa 8 hours after insemination. The most prominent effect of ageing of oocytes on early events of penetration was observed in the incidence of polyspermy rather than in the penetration rate and the proportion of pronuclear plus cleaved oocytes: the proportion of polyspermic oocytes significantly increased (P < 0.05) in oocytes inseminated after 28 to 48 hours of culture (36 to 78%) compared with those cultured for 20 to 24 hours (22 to 35%) for maturation. Culture experiments for early development of penetrated oocytes indicated that no significant differences were observed in the proportions of oocytes cleaved to the two- to four-cell stage 48 hours after insemination among those cultured for 20 to 40 hours for maturation. However, further cleavage to the four- to sixteen-cell stage 72 to 96 hours after insemination was greatly inhibited as ageing of oocytes proceeded from 28 hours in culture for maturation.