Effect of Fetal Serum and Follicular Liquor Supplementation on the In Vitro Production of Bovine Embryos

Abstract

To optimize In Vitro Fertilization (IVF) results in bovines by lowering costs, comparing the efficiency of Fetal Calf Serum (FCS) (high cost) with respect to bovine Follicular Fluid (bFF) (low cost) and by quantifying embryo production. Cumulus-Oocyte-Complexes (COC) obtained from a slaughter house, transported at 37°C in saline solution, were classified according to their quality in A, B, C and D. A and B oocytes were washed with modified Phosphate-Buffered Saline (m-PBS) and three groups were randomly formed (GC; G1; G2) cultured in drops of the respective media (10 COC/drop of 100 µL), covered with mineral oil and placed in an incubator (38,5°C, 5% CO₂ y 95% humidity). Maturation was done in Tissue Culture Medium (TCM-199) and antibiotics, for 22h and developed in CR1aa medium and antibiotics. The Control Group (CG) was supplemented during both maturation and development stages with 5% FCS and 10% bFF; G1 with 5% FCS during maturation and development and G2 with 10% bFF during maturation and development. We obtained bFF from follicles larger than 15 mm, it was centrifuged (800G) inactivating it. At fertilization Bracket and Oliphant (BO) medium was used. Zygotes were evaluated every 48 h for 8 days since insemination, counting the Division Rate (DR) and the Embryo Development (ED) of compacted morulae. χ² Test was applied. For RD, G2 differs from G1 and CG (p<0.05), CG and G1 do not differ significantly (p>0.05). During development no significant differences were found between groups (p>0.05). Results show that when using FCS, bFF or a combination of both, development results are similar. It is assumed that it is possible to substitute FCS with bFF.