title: In vitro embryo production in goats: Slaughterhouse and laparoscopic ovum pick up-derived oocytes have different kinetics and requirements regarding maturation media.
abstract: A total of 3427 goat oocytes were used in this study to identify possible differences during in vitro embryo production from slaughterhouse or laparoscopic ovum pick up (LOPU) oocytes. In experiment 1, one complex, one semi-defined, and one simplified IVM media were compared using slaughterhouse oocytes. In experiment 2, we checked the effect of oocyte origin (slaughterhouse or LOPU) on the kinetics of maturation (18 vs. 22 vs. 26 hours) when submitted to semi-defined or simplified media. In experiment 3, we determined the differences in embryo development between slaughterhouse and LOPU oocytes when submitted to both media and then to IVF or parthenogenetic activation (PA). Embryos from all groups were vitrified, and their viability evaluated in vitro after thawing. In experiment 1, no difference (P > 0.05) was detected among treatments for maturation rate (metaphase II [MII]; 88% on average), cleavage (72%), blastocyst from the initial number of cumulus oocyte complexes (46%) or from the cleaved ones (63%), hatching rate (69%), and the total number of blastomeres (187). In experiment 2, there was no difference of MII rate between slaughterhouse oocytes cultured for 18 or 22 hours, whereas the MII rate increased significantly (P < 0.05) between 18 and 22 hours for LOPU oocytes in the simplified medium. Moreover, slaughterhouse oocytes cultured in simplified medium matured significantly faster than LOPU oocytes at 18 and 22 hours (P < 0.05). In experiment 3, cleavage rate was significantly greater (P < 0.001) in all four groups of embryos produced by PA than IVF. Interestingly, PA reached similar rates for slaughterhouse oocytes cultured in both media, but improved (P < 0.05) the cleavage rate of LOPU oocytes. Slaughterhouse oocytes had acceptable cleavage rate after IVF (∼67%), whereas LOPU oocytes displayed a lower one (∼38%), in contrast to cleavage after PA. The percentage of blastocysts in relation to cleaved embryos was not affected by the origin of the oocytes (P > 0.05). Therefore, slaughterhouse oocytes developed a greater proportion of blastocysts than LOPU ones, expressed as the percentage of total cumulus oocyte complexes entering to IVM. Vitrified-thawed blastocysts presented similar survival and hatching rates between the oocyte origin, media, or method of activation. In conclusion, slaughterhouse and LOPU derived oocytes may have different IVM kinetics and require different IVM and IVF conditions. Although the IVM and IVF systems still need improvements to enhance embryo yield, the in vitro development step is able to generate good quality embryos from LOPU-derived oocytes.
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France; Laboratory of Physiology and Control of Reproduction, Veterinary School, State University of Ceará, Fortaleza, Ceara, Brazil. Electronic address: joannavet@gmail.com.
authors.forename: Joanna Maria G
authors.initials: JM
authors.lastname: Souza-Fabjan
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France; Museum National d'Histoire Naturelle, Réserve de la Haute Touche, Obterre, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Locatelli
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France; Museum National d'Histoire Naturelle, Réserve de la Haute Touche, Obterre, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Duffard
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Corbin
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Touzé
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Perreau
authors.affiliation: Laboratory of Endocrinology, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: Jean François
authors.initials: JF
authors.lastname: Beckers
authors.affiliation: Laboratory of Physiology and Control of Reproduction, Veterinary School, State University of Ceará, Fortaleza, Ceara, Brazil.
authors.forename: Vicente José F
authors.initials: VJ
authors.lastname: Freitas
authors.affiliation: INRA, UMR7247, Physiologie de la Reproduction et des Comportements, INRA, CNRS, Université de Tours, Haras Nationaux, Nouzilly, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Mermillod
mesh_terms: D000003:Abattoirs; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D018929:Cell Culture Techniques / Q000662:veterinary; D003470:Culture Media; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D006041:Goats / Q000196:embryology*; D010535:Laparoscopy / Q000662:veterinary*; D009865:Oocytes / Q000254:growth & development*; D010312:Parthenogenesis; D013396:Suction / Q000662:veterinary; D020858:Tissue and Organ Harvesting / Q000379:methods / Q000662:veterinary*
chemical_list: D003470:Culture Media
keywords: Blastocyst; Caprine; IVF; IVP; Laparoscopic ovum pick up; Oocyte maturation