Efeito do fator de crescimento epidermal (EGF) na maturação in vitro de oócitos caninos

Abstract

This work aimed to evaluate the influence of EGF on canine oocyte in vitro maturation (IVM). We carried out the transport of the ovaries in sodium chloride 0.9% solution, which were cut into phosphate-buffered saline (PBS) and the cumulus oocyte complexes (COCs) selected in tissue culture medium (TCM), supplemented with 199 HEPES. A total of 405 grade 1 oocytes were collected from bitches in anestrus and diestrus. Oocytes from the two phases of estrous cycle were subjected to two treatments: medium with addition of 10 ηg/mL epidermal growth factor (EGF) (T) and medium without supplementation (C). After 72 h of maturation, COCs were denuded, fixed and stained with Hoechst 33342 to assess nuclear maturation. The oocytes obtained from the ovaries of the diestrus phase of the T group demonstrated higher percentage (18.8%) of oocytes at metaphase II stage (M-II) than C group (1.3%) (p < 0.01). The oocytes from anestrus phase demonstrated difference (p < 0.05) between the groups, observing smaller proportion (4.1%) of oocytes at the stage of germinal vesicle (GV) in group T compared to group C (16.1%). Comparing the different reproductive status in media supplemented with EGF difference (p < 0.05) was observed only in oocytes diestrus phase relative to the stage of M-II. Thus, supplementation on maturation medium at a concentration of 10 ηg/mL of EGF in this study had positive influence only on IVM of canine oocytes obtained from diestrus phase, increasing the levels of M-II in this species.