The effects of lycopene and ascorbic acid on embryo development and quality supplement in vitro bovine embryo culture mediums

In this thesis study, the aim was to reduce oxidative stress and improve both the quantity and quality of embryos by supplementing the embryo culture medium with non-enzymatic antioxidants, Lycopene and Ascorbic acid, either separately or in combination. The material of the study consisted of 424 ovaries, which were transported to the laboratory under appropriate conditions from a slaughterhouse. Follicles with 2–8 mm diameters on the ovaries were aspirated to collect oocytes. The collected oocytes were classified based on their quality, and only A-grade immature oocytes were included in the study. Commercial solutions from the IVF Bioscience company were used for the maturation, fertilization, and culture processes of the oocytes. Oocyte maturation and fertilization were carried out in an incubator providing a temperature of 38.5°C, 5.5% CO₂, and a highly humid environment. The oocytes that completed maturation were subjected to the fertilization process. After the fertilization process was completed, the presumptive zygotes were transferred to the culture medium, and they were divided into groups with the addition of different doses of two separate antioxidants, both individually and in combination. The culture process was conducted in an incubator with 6% CO₂, 6% O₂, a temperature of 38.5°C, and high humidity for 6–9 days. Regarding fertilization as day 0, blastocysts in the IVC medium were examined starting from day 6. The differences in blastocyst numbers among the groups were statistically analyzed. Overall, it was statistically determined that the number of blastocysts and the blastocyst formation rates obtained from the combined groups were higher than those of the control and lycopene groups at different doses (P<0.05). However, no statistically significant difference was observed among the groups supplemented with lycopene and ascorbic acid (P>0.05). From the blastocysts obtained in the study, at least ten blastocysts per group were subjected to TUNEL staining. The number of cells in the blastocysts (inner cell mass, trophectoderm, and total cell numbers) and apoptotic cell counts were determined. The total cell numbers in the blastocysts were found to be significantly higher in the combined groups compared to the control and lycopene groups (P<0.05). When the number of apoptotic cells was examined, it was statistically determined that the lowest apoptotic cell count was found in the lycopene 0.2 µM and ascorbic acid 2 mM (Lik0.2+AA2) group (P<0.05). In conclusion, the addition of some antioxidants to the culture medium, either alone or in combination such as lycopene and ascorbic acid had a positive effect on development rates