Expressão gênica e viabilidade de folículos ovarianos pré-antrais de Sapajus apella congelados e cultivados in vitro

Abstract

The aim of the present study was to develop a freezing protocol for the preservation of preantral follicles from Sapajus apella (capuchin monkeys). To this end, ovarian fragments were exposed to different cryoprotectant solutions, added or not by antioxidants (selenium or trolox), frozen and in vitro cultived by 24 hours. Morphology, ultrastructure, viability, oxidative stress and mocelular analyses were performed. The collection site was in the Primates Nacional Center (CENP) and nine healthy mature female capuchin monkeys were used. Ovarian biopsies of 1 mm3 were collected by laparoscopy. The follicles were classified accordingly to their developmental phase in primordial, primary or secondary. Follicular viability scored using fluorescent markers (propidium iodide and Hoechst). qRT-PCR was used to evaluate the expression of hormones and growth factors. TEAC was used to measure oxidative stress in the tissue. In cryoprotectant solution containing trolox did not affected follicular morphology and gene expression. Cryopreservation resulted in higher rates of follicular viability when trolox was present in the solution, but expression of genes encoding BMP4 and KL was negatively affected. Our findings show a favorable effect of adding trolox to a cryopreservation solution. However, follicular viability and gene expression was affected after in vitro culture.