Evaluation of follicular genes pattern and growth of preantral follicles after culture in alginate hydrogel following vitrification of the mouse ovarian tissue

Asgari F.; Valojerdi M. R.; Ebrahimi B.
April 2015
Iranian Journal of Reproductive Medicine;Apr2015 Supplement, p73
Academic Journal
Introduction: This study was set up to evaluate the effect of ovarian tissue vitrification on the in vitro growth and pattern of follicular genes expression in mouse preantral follicles encapsulated within alginate hydrogel. Materials and Methods: Ovaries of 12-14 days old female NMRI mice allocated into fresh control and vitrification groups. For cryopreservation, ovaries equilibrated with a solution (ES) that composed of 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 15 min and vitrification was performed by a solution (VS) that composed of 15% EG and 15% DMSO for 30 min then ovaries loaded to nitrogen with needle. Descending concentrations of sucrose (1, 0.5, 0.25 M) used for warming step. After histologic assessment of ovaries, in the next stage, pre-antral follicles was mechanically isolated from control and vitrified ovaries and was cultured for 12 days in 0.7% sodium alginate. Preantral follicles survival rate, growth, antrum formation and relative expression of oocyte- specific genes (Bmp 15, Gdf9'Fgf8, Igfl, Kit, Kit ligand) was assessed after 1, 8 and 12 days of culture and finally maturation rate of oocytes was studied. Results: Preantral follicles in vitrified group showed a lower survival rate on 8 and 12 days of culture (p<0.05) but could retain a comparable morphological appearance, growth and antral formation with the control group. Reduction of Bmpl5, Gdf9, FgfS, Kit, Kit-l showed during 12 days of culture (p<0.05). Although the expression of Gd/9, Kit, Kit-l in vitrification group was more than control group at 1st day of culture but all genes in both groups showed same expression after 12 days of culture (p<0.05). Conclusion: Although vitrification of ovarian tissue reduces the survival rate, it is a safe method for preservation of preantral follicles and could not modify the relative expression of follicullar genes and oocytes maturation capacity.


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