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Manipulation of Follicular Development to Produce Developmentally Competent Bovine Oocytes¹

^a L'Alliance Boviteq, Inc., St-Hyacinthe, Quebec, Canada J2S 7A9 ^b IVF Labs, LLC, Salt Lake City, Utah 84117 ^c Faculté Des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Université Laval, Ste-Foy, Québec, Canada G1K 7P4

Superstimulation in donor cows increases the number of cumulus-oocyte complexes (COC), but when compared to in vivo maturation, in vitro maturation results in only half as many blastocysts after prolonged in vitro culture. The objective of this study was to establish a superstimulation protocol that would produce a maximal number of competent COC for standard in vitro embryo production. During experiment 1, eight cyclic Holstein heifers were superstimulated with four doses of FSH. Half the heifers received an injection of LH 6 h before ovum pick-up (OPU). The COC were collected following OPU either 33 or 48 h following the last FSH injection (coasting period). During experiment 2, six cyclic Holstein heifers were superstimulated with six doses of FSH, and in half the heifers, LH was administered 6 h before OPU. The COC were collected following ultrasound-guided transvaginal aspiration of both ovaries 48 h after the last FSH injection (coasting period). The COC originating from follicles with a diameter of 5 mm or more (n = 180 for experiment 1 and 57 for experiment 2) were subjected to standard in vitro maturation, fertilization, and development. When animals were administered four doses of FSH, 48 h of coasting resulted in significantly more 5- to 10-mm follicles (P < 0.01) than 33 h of coasting. If a 33-h coasting period was used, administration of LH 6 h before OPU resulted in a significant increase in both percentage of blastocysts and embryo production rate at Days 7 and 8 (P 0.05) of in vitro culture. If a 48-h coasting period was used, LH injection did not affect the rates of blastocyst production. When donors were administered six doses of FSH with a 48-h coasting period, the highest results, although not significant (P < 0.08), were obtained when animals received LH 6 h before OPU, with 80% ± 9% (mean ± SEM) blastocysts and 0.8 ± 0.09 embryo produced per COC retrieved per heifer at Day 8 of culture. Never has in vitro technology been so close to producing 100% developmentally competent COC.

First decision: 29 March 2001.

¹ Supported by NRC (IRAP), SEMEX Canada, NSERC, and Pacific Fertility Medical Centers.

² Correspondence. FAX: 418 656 3766; marc-andre.sirard{at}crbr.ulaval.ca

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