RESUMO
The aim of this study was to evaluate the effect of leukemia inhibitory factor (LIF) on the activation and survival of preantral follicles cultured in vitro enclosed in ovarian fragments (in situ). Goat ovarian cortex was divided into fragments to be used in this study. One fragment was immediately fixed (fresh control - FC) and the remaining fragments were cultured in supplemented minimum essential medium (MEM) without (cultured control - CC) or with different concentrations of LIF (1, 10, 50, 100 or 200 ng/ml) for 1 or 7 days, at 39°C in air with 5% CO2. Fresh control, CC and treated ovarian fragments were processed for histological and fluorescence analysis. The percentage of histological normal preantral follicles cultured for 7 days with 1 ng/ml (49.3%), 10 ng/ml (58.6%) and 50 ng/ml (58%) of LIF was higher than in the CC (32.6%; p < 0.05). After 7 days of culture, the percentage of primordial follicles in situ cultured with LIF decreased and primary follicles increased in all LIF concentrations compared with FC and CC (p < 0.05). In conclusion, LIF induced primordial follicle activation and supported preantral follicle viability of goat ovarian tissues cultured for 7 days.
Assuntos
Cabras/fisiologia , Fator Inibidor de Leucemia/farmacologia , Folículo Ovariano/efeitos dos fármacos , Técnicas de Cultura de Tecidos/veterinária , Animais , Sobrevivência Celular , Meios de Cultura/metabolismo , Feminino , Fluorescência , Cabras/anatomia & histologia , Cabras/metabolismo , Modelos Animais , Oócitos/citologia , Oócitos/metabolismo , Oócitos/fisiologia , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/metabolismo , Folículo Ovariano/fisiologia , Fatores de TempoRESUMO
The aim of this study was to evaluate the effects of pituitary (pFSH) or recombinant (rFSH) FSH on the survival and growth of caprine preantral follicles. Caprine ovarian tissues were in vitro cultured for one or seven days in Minimum Essential Medium (MEM) alone or containing 10, 50, 100 and 1000 ng/ml of pFSH or rFSH. Control tissues (non-cultured) and those cultured were processed for histological and ultrastructural studies. In addition, follicular and oocyte diameter were analysed. After seven days of culture, only 50ng/ml of rFSH maintained the percentage of normal follicles similar to control. Moreover, 10 ng/ml of pFSH and all the concentrations of rFSH promoted primordial follicles activation. In addition, the presence of 50 ng/ml of rFSH promoted the highest follicular diameter at day seven of culture. In conclusion, 50 ng/ml of rFSH maintained the ultrastructural integrity of caprine preantral follicles, promoted primordial follicles activation and further growth of cultured follicles.
O objetivo deste estudo foi avaliar os efeitos do FSH pituitário (pFSH)ou recombinante (rFSH) sobre a sobrevivência e o crescimento de folículos pré-antrais caprinos. O tecido ovariano foi cultivado in vitro por um ou sete dias em Meio Essencial Mínimo (MEM) sozinho, ou contendo 10, 50, 100 e 1000 ng/ml de pFSH ou rFSH. O grupo controle (não cultivado) e aqueles cultivados foram processados para análises histológica e ultra-estrutural. Além disso, os diâmetros folicular e oocitário foram avaliados. Após sete dias de cultivo, apenas 50 ng/ml de rFSH manteve o percentual de folículos normais semelhante ao controle. Além disso, 10 ng/ml de pFSH e todas as concentrações de rFSH promoveram ativação de folículos primordiais. A presença de50 ng/ml de rFSH promoveu o maior diâmetro folicular após sete dias de cultivo. Em conclusão, 50 ng/ml de rFSH manteve a integridadede folículos pré-antrais caprinos e promoveu a ativação e o crescimento dos folículos cultivados.
