| Autor: |
Leal ÉSS; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Vieira LA; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Sá NAR; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Silva GM; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Lunardi FO; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Ferreira ACA; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Campello CC; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Alves BG; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Cibin FWS; University Federal of Pampa, Uruguaiana-Rio Grande do Sul, Av. General Osório, 900 - São Jorge Bagé, RS - CE - 96400-100, Brazil., Smitz J; Follicle Biology Laboratory, Center for Reproductive Medicine, UZ Brussel, Laarbeeklaan 101, B-1090 Brussels, Belgium., Figueiredo JR; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil., Rodrigues APR; Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceará, Av. Dr Silas Munguba, 1700 - Campus of Itaperi, Fortaleza - CE - CEP 60741-903, Brazil. |
| Abstrakt: |
The aim of this study was to evaluate the viability, antrum formation and in vitro development of isolated secondary follicles from vitrified caprine ovarian cortex in a medium previously established for fresh isolated secondary follicles, in the absence (α-minimum essential medium (α-MEM+) alone) or presence of FSH and vascular endothelial growth factor (VEGF; α-MEM++FSH+VEGF). Ovarian fragments were distributed among five treatments (T1 to T5): fresh follicles were fixed immediately (T1), follicles from fresh tissue were cultured in vitro in α-MEM+ (T2) or α-MEM++FSH+VEGF (T3) and follicles from vitrified tissue were cultured in vitro in α-MEM+ (T4) or α-MEM++FSH+VEGF (T5). After 6 days of culture, treated follicles (T2, T3, T4 and T5) were evaluated for morphology, viability and follicular development (growth, antrum formation and proliferation of granulosa cells by Ki67 and argyrophilic nucleolar organiser region (AgNOR) staining). The levels of reactive oxygen species (ROS) in the culture media were also assessed. Overall, morphology of vitrified follicles was altered (P<0.05) compared with the fresh follicles. Follicular viability, antrum formation and ROS were similar between treatments (P>0.05). The average overall and daily follicular growth was highest (P<0.05) in T3. Granulosa cells in all treatments (T1, T2, T3, T4 and T5) stained positive for Ki67. However, fresh follicles from T3 had significantly higher AgNOR staining (P<0.05) compared with follicles of T1, T2, T4 and T5. In conclusion, secondary follicles can be isolated from vitrified and warmed ovarian cortex and survive and form an antrum when growing in an in vitro culture for 6 days. |
