Utilização do ácido ascórbico na produção in vitro de embriões bovinos

Abstract

The production of embryos in vitro (PIVE) has become a well-established technique. But the search for tools has been tested to improve results. Among them, the use of ascorbic acid that has become widely studied by researchers, because its function may be to reduce oxidative stress, which today is a reality of PIVE. The objective of this work was to evaluate the effect of ascorbic acid on PIVE, from maturation to hatching of blastocysts. The experiment was carried out at the Animal Reproduction Biotechnology Laboratory (BIOTEC) of the State University of Goiás. The ovaries came from a local slaughterhouse and transported to the laboratory where they were washed and then aspirated. Only grade I and II oocytes were selected for use in PIVE. The oocytes were divided into AAMIV, AAMC and AACIV groups, and among the groups was used ascorbic acid in three concentrations (10 μg / mL, 50 μg / mL and 100 μg / mL) plus control (0 μg / mL ascorbic acid). submitted to maturation for 24 hours with medium supplemented with ascorbic acid in the AAMIV and AAMC group. In vitro fertilization was used semen from a single bull and broken, with known fertility, and from central. Possible zygotes were cultivated for 11 days. In cultivation, ascorbic acid was added to the AACIV and AAMC group. All steps were performed in a cell culture greenhouse at 38.8ºC and 5.5% CO2 gas atmosphere. Statistical analyzes were performed by ANOVA and Bonferroni analysis at 5% significance. The rate of blastocysts in relation to the cleaved ones (AAMIV², AAMC² and AACIV²) differed (p <0,05) from each other. Therefore, the concentrations of 10 μg / mL and 50 μg / mL of the AAMIV² group were the ones that presented the best results (65.23% and 54.50%), comparing between the groups with ascorbic acid. Comparing the cleavage rate in the other ascorbic acid concentrations of the groups, AAMC¹ at a concentration of 10 μg / mL (84.14%), 50 μg / mL (82.68%), and 100 μg / mL (84, 16%), they differ (p <0.05) from each other. The blastocyst hatching rate at D11 of culture did not show statistical difference when compared between groups. Finally, further studies on the antioxidant have yet to be done.