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ADDITION OF VITAMIN E IN THE MIDDLE OF VITRIFICATION VIABILITY AND DEVELOPMENT IN VITRO OF OOCYTES GATAS DOMESTIC POST-THAW

Grant number: 12/01580-8
Support type:Scholarships in Brazil - Master
Effective date (Start): May 01, 2012
Effective date (End): April 30, 2014
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal researcher:Wilter Ricardo Russiano Vicente
Grantee:Raquel Ribeiro Gutierrez
Home Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The use of gamete cryopreservation techniques in domestic cats is an important method in improving assisted reproduction procedures. Among these, vitrification is rapid procedure, practical and based in the use of high concentrations of cryoprotectants with the intention to inhibit the formation of ice crystals. Objectively evaluate whether the antioxidant addition (vitamin E) the protocols for vitrification of oocytes of gatas domestic increases the viability oocyte post-thawing, incrementing rates in vitro maturation. For this, we conducted three experiments. In experiment I will be compared two protocols of vitrification using as base medium (MB) the TCM199 with sucrose plus cryoprotectants: MB with EG + Me2SO (ethylene glycol and dimethylsulfoxide) and MB with EG + PrOH (ethylene glycol and propylene glycol); 70 oocytes per group. After thawing, this material will be evaluated for morphological changes and viability staining cFDA / trypan blue. In experiment II will be employed the protocol of experiment I with the best score, plus three concentrations of vitamin E (0.4, 0.6 and 0.8 mmol) with equal numbers of oocytes per concentration (group), and after thawing are evaluated for morphological changes and viability. Experiment II oocytes that maintain their morphology after thawing will undergo in vitro maturation for a period of 36 hours (experiment III). The evaluation of their development stage meiotic well as the dispersion of cortical granules post IVM will be made with the coloration FITC-LCA-Hoescht 33 342. The percentage of oocytes considered viable after vitrification and those that resumed meiosis after in vitro maturation, will be analyzed by the Kruskal-Wallis test (p <0.05).

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