P-219: GDF9, BMP15 and Their Receptors Expression During In Vitro Culture of Mouse Vitrified Ovarian Tissue Derived Preantral Follicles
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Abstract:
Background: In vitro culture (IVC) of isolated preantral follicles from cryopreserved ovarian tissue might be an efficient method for enhancing mature oocytes in patients who are exposed to infertility. Materials and Methods: Ovaries of 13-day old NMRI mice were removed and randomly placed into control,needle immersed (NIV) and solid surface vitrification (SSV) groups. For vitrification, ovaries were transferred into equilibration [7.5% (EG and DMSO)] and vitrification medium [15% (EG and DMSO) and 0.5 M sucrose], then they immersed in liquid nitrogen after loading by acupuncture needle in NIV group and cooling on pre cooled steel surface in SSV group. Thawing was done in 2steps (1 and 0 M sucrose solution). Mechanically isolated preantral follicles were cultured in α-MEM supplemented with (FSH, LH, ITS, FBS) for 12 days. The expression rate of maturation genes (GDF9 and BMP15) and their receptors (BMPR2, ALK5 and ALK6) in all experimental groups were evaluated quantitatively by real time PCR after 24 hour, 6, 10 and 12 days of culture. Results: No significant difference was observed in the expression of maturation genes and their receptors between vitrification (NIV and SSV) and control groups and also between NIV and SSV groups. It must be noted that the expression patterns of mentioned genes in two vitrification groups were similar to the control one and assimilate the in vivo pattern in somehow. Conclusion: Although the cooling rate in SSV method is delayed compared to NIV method, the pattern of maturation genes expression during IVC was similar in both groups and it seems that preantral follicles compensate cryoinjuries during IVC period.
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Journal title
volume 6 issue 2
pages -
publication date 2012-09-01
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