UNIVERSIDADE ESTADUAL DO CEARÁ FACULDADE DE VETERINÁRIA Programa de Pós-Graduação em Ciências Veterinárias

The objectives of this thesis were: 1) To determine the effect of transport temperature (4 vs 33oC) on morphology, follicular growth, viability and oocyte maturation, and production of reactive oxygen species (ROS) of preantral and early antral follicles goats for 18 days in vitro culture (Phase I); 2) to evaluate follicular survival and activation rates after 7 days of in vitro culture in a three-dimensional system in the absence (isolated primordial follicles and included in Alginate 0.8%) or in the presence of ovarian tissue (Phase II); 3) To investigate the survival, activation and follicular growth after 7 days of in vitro culture of ovarian fragments of baboons (Phase III) included in different matrices (Alginate 1% or poly(ethylene glycol)-vinyl sulfone (PEG) 5%); 4) Investigate the effect of different concentrations of Cilostamida (10 and 20 μM) and exposure times (6 and 12 hours) during IVF of bovine and goat oocytes (Phase IV). In Phase I, goat ovaries were individually transported in MEM-HEPES at 4 or 33 ° C. Subsequently, the follicles were cultured in vitro for 18 days and cumulus-oocyte (CCO) complexes matured for 32 hours. In Phase II, the primordial goat follicles included in Alginate (10 follicles / drop), or wrapped in ovarian tissue (in situ) encapsulated or not in Alginate, were cultured for 7 days. In Phase III, precryopreserved baboon follicles were cultured for 10 days in ovarian tissue (in situ), in the presence or absence of extracellular matrices (PEG-VS vs. Alginate) and FSH. Finally, in the Phase IV, the goat and cattle COCs were cultured for 30 hours, starting with in vitro prematuration (IVPM) for 6 or 12 hours before IVM (24 or 18h). The best result for the production of fully-grown oocytes and meiotic resumption was obtained using low (4 ° C) for preantral follicles and high (33 ° C) for initial antral follicles (Phase I). Phase II results showed that caprine primordial follicles isolated and encapsulated in Alginate presented satisfactory rates of survival and higher percentages of follicular activation when compared to the other treatments. In the Phase III, treatments containing medium supplemented with FSH associated and extracellular matrices (Alginate and PEG-VS) promoted an improvement in the survival of baboon preantral follicles included in the ovarian tissue after 10 days in vitro culture. In the Phase IV, the combination of concentration/exposure time to cilostamide during IVPM delayed the meiotic progression only in the bovine species after 6 and 12 hours of culture. In conclusion, the transport temperature differentially affected the in vitro development of preantral follicles and initial antral follicles (Phase I); the in vitro culture of isolated goat primordial follicles enclosed in Alginate allowed a better rate of follicular activation (Phase II); In baboons, the use of extracellular matrices in the presence of FSH improved follicular survival after 10 days of in vitro culture (Phase III). Finally, in the Phase IV, the cilostamide delayed in a concentration/dependent manner the meiotic resumption in bovine species only.