The cumulus cells reflect the succes of IVF of porcine oocytes treated with dimethylthiourea

LOMBARDO, DM.; LORENZO, MS.; CRUZANS, PR.; TEPLITZ, G.; TELLO, MF.

Bangkok

Conferencia; 44th IETS Annual Conference; 2018

International Embryo Technology Society

The aim of this study was to evaluate the effect of the antioxidant dimethylthiourea (DMTU) as a supplement during the collection and washing of cumulus oocyte complexes (COC) on the apoptosis in cumulus cells (CC) and the competition for oocyte development in vitro. COC were obtained by follicular aspiration. The aspiration and washing medium in the control were TCM 199 with 10% porcine follicular fluid, 0.3 mM of sodium pyruvate and penicillin/streptomycin. In the treatments, 2 mM of DMTU or 20 mM of DMTU were added. After 44 h of in vitro maturation (IVM) the percentage of nuclear maturation was determined by staining of the denuded oocytes with Hoechst 33442 (n = 357). After 18 h of in vitro fertilization (IVF) with 15°C refrigerated spermatozoa from boars with proven fertility, the suspected zygotes were stained with Hoechst 33342 to evaluate the percentages of sperm penetration (SP), monospermic penetration (MP), male pronucleus formation (MPN) and IVF efficiency (monospermic oocytes/total oocytes) (n = 260). In post-MIV CC, flow cytometry was performed after staining with Annexin V-Propidium Iodide to evaluate apoptosis and viability (n = 90000). Data were analysed with Flowing V 2.5.1. A comparison of proportion among categories was used, significant differences were determined with p