Abstract
ObjectivesTo evaluate the effects of ablation of steroid production and the role of testosterone and dihydrotestosterone in the development of secondary follicles of nonhuman primates (Macaca mulatta) in 3D matrix. MethodsThe ovarian cortex (n=8 adult females) was diced and secondary follicles were isolated and cultured individually for 40 days on alginate matrix. Experiment 1: Ablation of steroid production with use of trilostane (TRL) and testosterone replacement. Follicles of 4 animals were assigned to 4 groups: control (vehicle), TRL (TRL from the start of culture), Low testosterone (T) (TRL testosterone and 10ng/mL), high T (TRL testosterone and 50ng/mL); Experiment 2: Ablation of steroid production with use of trilostane (TRL) and replacement of dihydrotestosterone (DHT). Follicles of 4 animals were assigned to 4 groups: control (vehicle), TRL (TRL from the start of culture), DHT (DHT 50ng/mL), DHT+TRL (TRL and DHT 50ng/mL). It were analyzed: survival, growth, antrum formation, steroid hormone production, and degree of maturation and quality of oocytes. ResultsThe percentage of surviving follicles in culture, growth and antrum formation was lower in the presence of TRL in all experiments, compared with the control group. Replacement of T and DHT in the presence of TRL, recovered survival, growth, antrum formation, hormone production, and the quality and maturity of oocytes comparable to the control group. DHT added to standard maturation media promoted an increase in the percentage of survival follicles and antrum formation compared to the control group. ConclusionsThis study provided evidence of the role of androgens since the beginning of folliculogenesis in primates, reinforcing the existence of molecular mechanisms regulating androgenic activity, and new interactions between androgens and steroid hormones. Our findings show that T and DHT can recover survival, growth, antrum formation, hormone production and oocyte viability of in vitro cultured preantral follicles exposed to ablation of steroid production. DHT added to standard maturation media promoted an increase in the percentage of follicle survival and antrum formation compared to the control group, opening perspectives to evaluate whether this androgen supplementation could improve follicular development in vitro. The knowledge obtained contribute to a understanding of the dynamics of folliculogenesis in primates, still poorly understood, and assists in identifying the optimal conditions for growing follicle, which may in future be applied to assist cancer patients who wish to preserve their future fertility
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