REACTIVE OXYGEN SPECIES PRODUCTION IN VITRIFIED PORCINE OOCYTES

DANIELA PINCHETTI; AILÉN APARICIO; PABLO CETICA; GABRIEL ÁLVAREZ; GABRIEL DALVIT; SERGIO MORADO

Jornada; XVIII Jornadas de la Sociedad Argentina de Biología; 2016

In order to evaluate reactive oxygen species (ROS) production in porcine oocytes matured in vitro and subjected to the process of vitrification-warming, Cumulus-oocyte complexes were obtained by aspiration of antral ovarian follicles and matured in medium 199 with porcine follicular fluid, FSH and LH, and incubated at 39°C, 5% CO2 in humidified air for 48h. Matured oocytes (n = 75) were denuded and a group was cryopreserved using the minimum volume vitrification method Cryotech® .Then oocytes were subjected to warning. ROS were measured at 0, 3 and 21h by the incubation of oocytes in PBS-PVA and DCHF-DA, while a sample was incubated to evaluate esterase activity in PBS-PVA and FDA. Fluorescence levels of DCHF-DA depends on intracellular esterase activity, so the ratio between the brightness obtained from DCHF-DA for each oocyte and the average brightness by FDA of each treatment was considered as a measure of ROS production. In fresh oocytes ROS production was higher at 0 compared with 3 and 21h (p