P-233: The Effect of Cumulus Cells in Biphasic In Vitro Maturation of Mice Germinal Vesicle Oocytes in The Presence of Alpha Lipoic Acid

Background: The efficiency of in vitro maturation (IVM) has remained low despite extensive effort to understand the culture conditions. This study investigated the effect of cumulus cells and meiotic inhibitors on oocyte developmental competence in the presence of Alpha Lipoic acid (ALA) Materials and Methods: Mouse germinal vesicle oocytes were considered in cumulus-denuded oocytes (CDOs) and cumulus-oocyte complexes (COCs) groups. GV oocytes from each main group were randomly distributed between two subgroups: IVM in the presence or absence of ALA. Each subgroups were divided into two groups as follow: (I) IVM without inhibitors only for 24 hours (control) and (II) IVM with 50uM Forskolin in combination with 10 uM Forskolin in two step manner: first step, GV oocytes were arrested meiotically in TCM199 containing Forskolin+Cilostamide and then, in the second step were matured without those for 24 hours. Obtained metaphase II oocytes were subjected to in vitro fertilization (IVF). Results: The COCs maturation, fertilization and two cells embryo rates were higher than CDOs in control group (p<0.05). Immature oocytes cultured in biphasic manner showed a significantly higher (p<0.05) maturation, fertilization and two cells embryo rates in COCs and CDOs compared to their control groups, while no significant difference was observed between relevant COCs and CDOs in the presence or absence of ALA (p>0.05) Conclusion: Supplemented maturation medium with Cilostamide+Forskolin could improve the developmental competence of GV oocytes. While, Adding ALA could not improve developmental competence. Meiotic inhibitors were more effective in CDOs than COCs