EFFECT OF IONOMYCIN ON THE IN VITRO FERTILIZATION OF PIGS

LUCHETTI, CG.; LORENZO, MS.; LOMBARDO, DM.

Ciudad Autónoma de Buenos Aires

Jornada; XX Jornada Anual de la Sociedad de Biología (SAB )2018; 2018

Sociedad Argentina de Biología

Given the relevance of porcine embryo production for commercial purposes or for biomedical studies, many research laboratories havefocused on the optimization of the in vitro fertilization (IVF). To improve its efficiency, the effects of ionomycin (Io) on IVF werestudied. Porcine COC obtained by follicular aspiration from slaughterhouse ovaries were in vitro matured in TCM-199 + porcinefollicular fluid (10%), FGF (3 ng / mL) and FSH (30 ug / mL) during 44-47 h. Then, matured oocytes were denuded with hyaluronidaseand treated with 5 uM ionomycin for 4 min and washed in TALP-H medium four times (group Io+), or without it: control (group C).Immediately, oocytes were subjected to IVF (coincubated 30 min with fresh boar semen 1x106 sperm / mL in modified BO medium). Thepresumptive zygotes were in vitro cultured in 50 ul drops of SOF medium at 39° C with 5% CO2, 5% O2 and 100% humidity. After 20 hform the beginning of the IVF, embryos were fixed in paraformaldehyde 4% during 30 min, washed and stored in PBS-PVA until use.To assess fertilization parameters presumptive zygotes were stained with Hoëchst and mounted for evaluation. The fertilizationparameters evaluated were: penetration (oocytes penetrated/ inseminated), masculine pronuclear formation (MPN=oocytes containingMPN/penetrated), monospermy (oocytes containing only one sperm head or one male pronucleus/ penetrated), and efficiency offertilization (monospermic oocytes/inseminated). Data were analyzed by Fisher´s exact test considering significant P