Laparoscopic ovum-pick up and in vitro embryo production in gonadotropin-stimulated gilts: Preliminary results and envisioned applications

The present study was conducted to establish if laparoscopic ovum pick-up (LOPU) could be adapted to the swine species, and if the developmental competence of LOPU-sourced oocytes from peripubertal gilts could be improved by gonadotropin stimulation, by comparing with oocytes sourced from slaughtered gilts lacking hormonal stimulation. Estrus was synchronized in 34 gilts of ∼6–8 months of age by daily oral administration of 17.6mg altrenogest for 13 days and 10 mg dinoprost IM on the last day of altrenogest. Follicular development was stimulated in all gilts with a single injection of 1250 IU eCG given 3 days before LOPU (together with the 12th dose of altrenogest). In about half of the gilts (Group eCG-hCG), 500 IU hCG were injected IM ∼72h after eCG injection, or ∼16–18h prior to LOPU, to initiate oocyte maturation in vivo, while the remaining animals only received eCG (Group eCG). Most gilts underwent LOPU twice alternating the gonadotropin protocol, thereby decreasing the impact of individual variation on results. Abattoir-sourced oocytes from prepubertal gilts served as Control. Following LOPU, oocytes were in vitro matured, fertilized, and cultured to the blastocyst stage following standard procedures, while oocytes collected from Group eCG-hCG gilts were considered partly matured in vivo and were matured for ∼24h instead of ∼44h. Embryos reaching the blastocyst stage were fixed and stained to assess quality through cell numbers. There were no significant differences in the number of follicles aspirated and cumulus-oocyte complexes (COC's) recovered between Groups eCG-hCG and eCG (22.4 and 16.9 vs. 22.6 and 17.6, P > 0.05), as well as the recovery rate (76.6 vs. 78.1, P > 0.05). Cleavage rate was not different between Group eCG-hCG, Group eCG and Control (61.1 vs. 64.4 vs. 53.4 %, P > 0.05). However, the blastocyst rate over total oocytes (32.2 vs. 36.9 vs. 11.1 %, P < 0.05), blastocyst rate over cleaved oocytes (51.8 vs. 55.1 vs. 21.2 %, P < 0.01) and the average number of cells/blastocyst (89.6 vs. 87.5 vs. 62.2, P < 0.01) were unaffected by hCG treatment in LOPU-sourced oocytes, but both LOPU groups were significantly higher than abattoir-sourced oocytes, respectively. Our results suggest LOPU may become a powerful tool for sourcing swine oocytes with higher developmental competence than abattoir-sourced oocytes and known disease status for creating swine models for human biomedical applications, as well as for accelerated genetic gain in swine breeding programs.