Viabilidad de ovocitos vitrificados y madurados in vitro de gata doméstica adulta (Felis catus) en estación reproductiva

Abstract

Vitrification of mammalian oocytes is a technique considered in experimentation. Oocyte survival is extremely variable, according to the techniques used and influenced by a number of conditions. The objective of this study was to determine the effects of adult domestic cat oocyte matured in vitro and vitrified in breeding season. We obtained 33 ovaries from housecats in good nutritional status without hormonal treatment, ovariectomized by a veterinary professional at home. The ovaries were transported to the laboratory and fragmented by microdissection under a stereoscopic microscope in a petri dish with modified buffer saline phosphate solution, inactivated calf serum and antibiotics to the collection, evaluation and selection of good quality cumulus-oocyte complexes (COCs). 506 COCs were vitrified in 0.25 ml straws each with 10–12 oocytes and stored for 30 days in liquid nitrogen (N2) at -196 °C. Then the straws were thawed, recovering 320 CCOs, which were matured in vitro. After this time the COCs were evaluated, discarding 50 which showed signs of degeneration. In the remaining 270 COCs good cumulus expansion was observed and also uniformly dark cytoplasm and integrity of the zona pellucida. The latter were divided into two similar groups to assess the feasibility; one was stained with Methylthiazblyl tetrazolium and the other with Trypan Blue. Both tested positive for the viability of the CCOs was found. The analysis of the results allow us to conclude that vitrification compromised the integrity of a large number of CCOs although more than 50% responded favorably in culture, showing signs of expected viability. According to these results it is necessary to further improve the protocol to increase the percentage of viability.