Effects of Chick Embryo Extract on Cx ۴۳, Cx ۳۷, Bmp۶ and, Gdf۹ Genes Expression In Vitro in Preantral Follicles Isolated from Vitrified and Non-Vitrified Mouse Whole Ovary

Background: Currently, vitrification of the ovarian tissue isused as an effective way to preserve oocytes and follicles. It appearsto be the case that chick embryo extract (CEE) can enablesatisfactory results in puberty of prenatal follicles.Materials and Methods: For this study, ovaries of ۱۰-۱۲ dayold Naval Medical Research Institute (NMRI) female micedivided into two groups: non-vitrified and vitrified. The preantralfollicles with a mean average diameter of ۱۱۰-۱۳۰ μmwere isolated by mechanical methods of non-vitrified and vitrifiedovaries and cultured in the aforementioned groups. First tosecond groups consisted of: non-vitrified ovaries in base solution(first group), base solution with ۵% chick embryo extract(second group), and third to fourth groups consisted of vitrified-warmed ovaries in base solution (third group) base solutionplus ۵% chick embryo extract (fourth group). The evaluationsincluded the survival rate of the follicles, the antrum formationrate, and maturation rate of oocyte that reached the metaphasetwo stage. Estradiol hormone levels, survival of follicles, morphologyof oocytes, and ovarian tissue histology were evaluated.Also, quantitative expression of oocyte maturation genes(Cx۴۳, Cx۳۷, Bmp۶, and Gdf۹) was evaluated after ۲۴ hoursand ۱۲ days of culture.Results: The amount of antrum formation was significantlyhigher in the ۵% chick embryo extract in vitrified and non-vitrifiedgroups than other groups (P<۰.۰۵). The Connexin ۴۳ genein vitro cultured group, with ۵% chick embryo extract, showedan analytically higher expression than the control group (vitrified,P<۰.۰۵ ).Conclusion: Considering the significant difference in the antrumformation and gene expression of Connexin ۴۳ betweenthe CEE and control groups, it seems that CEE could improvethe quality and quantity of matured follicles in vitro but the,other ovarian functional factors assays, including fertilizationability and developmental potential of derived embryos are necessary.

کلیدواژه ها:

Preantral Follicle ، Chick Embryo Extract ، Ovarian Tissue ، Vitrification ، In Vitro