Effects of Chick Embryo Extract on Gdf۹, Bmp۶, Cx۳۷, Cx۴۳ Genes Expression in Non-Vitrified and Vitrified Mouse Preantral Follicles during In Vitro Culture

Background: Vitrification and in vitro culture of preantral folliclesare a suitable way to female's fertility preservation thatare exposed to the infertility risks. The development of preantralfollicles culture system can increase the growth rate andquality of oocytes. To this aim, the effects of various biofactorssuch as proteins, sugars and other factors have been investigated.Chick embryo extract (CEE) contains various factors suchas growth factors, vitamins, mineral, sugar, protein and lipids,can stimulate growth and differentiation of the cells. In this research,the effects of chick embryo extract on mouse preantralfollicles in vitro culture has been investigated.Materials and Methods: For this study, preantral follicles witha diameter of ۱۱۰-۱۳۰ μm were isolated from ۱۲-۱۴ days oldfemale mouse and divided and in vitro cultured ۱۲ days into twogroups of Fresh (non-vitrified) and Vitrified. The base culturemedium (BM) contains: α-MEM + ۵% FBS + ۱% ITS (۵mg /ml) + ۱% FSH. Each vitrified and non-vitrified follicles wereclassified into ۴ groups: ۱. Fresh BM group: culture of nonvitrifiedpreantral follicles in the base medium ۲. Fresh CEEgroup: culture of non-vitrified preantral follicles were in a basemedium of + ۵% chick embryo extract ۳. Vit BM group: culture of vitrified-warmed preantral follicles in the base medium and۴. Vit CEE group: culture of vitrified-warmed preantral follicles,in base medium + ۵% chick embryo extract. The follicleswere cultured for ۱۲ days after melting in sucrose solutionswith decreasing concentrations. Survival rates were measuredon days ۱, ۴, ۸ and ۱۲ after culture, as well as expression ofGdf۹, Bmp۶, Cx۴۳ and Cx۳۷ genes on the ۱۲th day after culturein all groups.Results: The results of this study showed the lowest survival offollicles during different days of culture in Vit BM group, comparedto Fresh CEE group (P<۰.۰۵). Antrum formation in VitCEE and Fresh CEE groups was significantly higher than VitBM and Fresh BM groups respectively (P<۰.۰۵).The expressionof Cx۴۳ gene on the ۱۲th day in the Vit CEE group wassignificantly higher than Vit BM group(P<۰.۰۵).Conclusion: Therefore, chick embryo extract as a natural nutritionalsupplement can be used in vitrification and in vitro cultureof mouse preantral follicles in future study.

کلیدواژه ها:

Chick Embryo Extract ، Preantral Follicle ، In Vitro Culture ، Vitrification