P-154: Comparison of Ovine Matured Oocytes Quality in SPOM Medium Supplemented with Different Doses of Oocyte-Secreted Factors

Background: In this study we evaluate the effects of different doses of GDF9 with fixed dose of BMP15 on quality of in vitro maturated sheep oocytes. Materials and Methods: Good ovine Cumulus-oocyte complexes (COCs) were first transferred to Pre-IVM medium, and then to three IVM mediums supplemented with 10%v/v BMP15 and: 1) 175 ng/ml 2) 131 ng/ml or 3) 219 ng/ml GDF9. After maturation period, MII oocytes with normal first polar body were counted. Oocyte viability was determined by means of the TUNEL technique. Immunofluorescence microscopy staining were used to evaluate cell meiotic spindle and redistribution of cortical granules (CGs). In addition, fertility rate in group 1 and 2 was determined by means of IVF. Statistical analyses were carried out by ANOVA and significant differences between means were determined using Tukey HSD test for comparison of multiple means. Results: Maturation rate of group 3 was significantly lower than group 1 and 2 (75.57% ± 2.74 vs. 93/56% ± 0.56 and 90.67% ± 4.09, respectively, p<0.05). Spindle,CGs, and TUNEL staining results indicated no significant difference between groups. MII oocytes with normal polar body in group 1 was significantly higher than group 2 and 3 (71.39% ± 0.75 vs. 25.80% ± 1.09 and 31.68% ± 2.36, respectively, p<0.001). Finally, the blastocyst formation in group 1 was significantly higher than group 2 (39.86% ± 0.38 vs. 28.73% ± 1.77, p<0.01). Conclusion: These results demonstrate that SPOM medium supplemented with 175 ng/ml GDF9 and 10% BMP15 enhance oocyte developmental competence.