Obtaining bovine cattle embryos in vitro at different time parameters of the activation onset of in vitro oocyte maturation
Abstract
Aim. To investigate the effect of using different time parameters of starting in vitro activation of oocyte maturation (activation onset) on the viability and further development of oocyte-cumulus complexes (OCC) of cows, the efficiency of oocyte fertilization, and the production and further in vitro development of embryos. Methods. Biotechnological, embryological, morphological, and cytogenetic methods, as well as methods of variation statistics were used in the research. The ovaries of the Ukrainian black-and-white dairy cows (n=28) were kept at the temperature of +38°C in the physiological solution. The OCC were obtained by incising visible antral follicles with a blade. The period from the moment of incising the antral follicles to placing the OCC for cultivation was 30-40 min in all the groups. The embryos with more than 16 cells were classified as morulae and the embryos with blastocoel as blastocysts. All the experimental groups were analyzed in three repeats (n=3). The statistical processing of the data was conducted using the reliability criterion x^2, and the differences were deemed statistically significant at p<0.05. Results. The study was conducted from 2021 to 2024. The effect of the activation procedure and different terms of its start on the biological completeness and further development of cow oocytes of Ukrainian black-and-white dairy breed (n-28) after meiotic maturation in vitro was studied. The coefficient and index of the fragmentation of the obtained in vitro embryos were analyzed for different time parameters of the activation onset. It was shown that the efficiency of obtaining bovine cattle embryos in vitro and their further development depended on the time of activation onset. The least number of gametes at the metaphase-2 stage (48.8%) and the most cells with degenerated chromatin (44.0%) were obtained when using the most delayed time of activation onset of in vitro maturation of oocytes (≥ 8 hours). In case of activation 2, 4, 6, and 8 hours after the start of cultivation, the rate of morula-blastocyst formation was 11.1, 9.0, 2.9, and 0.0%, respectively. Conclusions. With an increase in the time interval between the start of culture and the activation of oocytes up to 8 hours, the number of the formed in vitro bovine embryos decreased.
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