P-88: Expression Pattern of Maturation Genes During In Vitro Culture of Alginate Encapsulated Preantral Follicles Derived From Frozen-Thawed Mouse Ovaries
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Abstract:
Background: This study was set up to evaluate the effect of ovarian tissue slow freezing on in vitro growth and pattern of maturation genes expression in mouse preantral follicles encapsulated within alginate hydrogel. Materials and Methods: Ovaries of 12-14 days old female NMRI mice were randomly allocated into control and slow freezing groups. In slow freezing group, ovaries were equilibrated in the cryoprotectant solution using dimethyl sulfoxide (DMSO), and then were cooled in a programmable freezer according to Min Xu et al., 2009. Ovaries were thawed in a stepwise manner and their morphology was studied histologically (HandE staining). Then in both control and experimental frozen- thawed groups, pre-antral follicles were mechanically isolated from ovaries and cultured for 12 days in 0.7% alginate hydrogels. Survival rate, follicular growth, antrum formation and relative expression of maturation genes (Bmp15, Gdf9, Fgf8, Igf1, Kit, Kit-l) was assessed after 1, 8 and 12 days of culture and finally maturation rate of oocytes was studied. Results: Morphological integrity of ovarian tissue was similar in both groups. However after in vitro culture significant lower survival and antrum formation rates were observed in slow freezing group compared to the control one (p<0.05). At the end of culture period, no significant difference reported in oocyte maturation rate (p>0.05). After 12 days of culture, Preantral follicles in cryopreserved group showed similar pattern of maturation genes expression compared to the control group. The pattern of Bmp15, Gdf9, Fgf8, Kit and Kit-l were down regulated during culture. Also, the expression of Kit and Kit-l in slow freezing group was lower than control group at day 8 of culture. Conclusion: Although slow freezing of ovarian tissue reduces the survival and antrum formation rate of encapsulated cultured preantral follicles, it could not modify the expression pattern of maturation genes and oocytes maturation capacity.
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Journal title
volume 8 issue 2.5
pages 103- 103
publication date 2014-07-01
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