Evaluation the expression of genes related to angiogenesis after vitrified ovarian tissue

Background: Many attempts were done for the preservation of fertility in cancer patients by improvement of ovarian tissue vitrification and transplantation. This study investigated the effect of direct cover vitrification (DCV) on the expression of angiogenic factors in vitrified ovarian tissue grafts.Methods: All phases of current experiment were in accordance to published guideline of The Care and Use of Laboratory Animals [NIH Publication No. 85-23, revised 1996] and approved by the local ethical committee of Tabriz University of Medical Sciences. Ovarian tissue was dissected from 6 to 8-week-old female Balb/c mice under anesthesia and the right ovaries immediately washed in α-MEM and left ovaries vitrified/thawed by direct cover vitrification (DCV2 and DCV3) cryoprotectant. Then ovaries were autotransplanted subcutaneously. After 7 days, tissues were dissected and assessed for expression of angiogenic factors such as vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) by real-time PCR.Result: We demonstrated for the first time the expression of VEGF and Ang2 in cryopreserved ovarian tissue graft by direct cover vitrification. In vitrified/grafted ovarian tissues, we observed the highest expression of VEGF and Ang-2 was observed in as the DCV2 group than DCV3 group.Conclusion: These findings suggest that DCV2 is a more efficient vitrification solution for expression of angiogenic factors after ovarian tissue transplantation. However, further investigations are necessary to optimize vitrification technique in order to improve revascularization in the early stage of transplantation.

Keywords:

Ang-2 , Real-time PCR , VEGF , Angiogenesis , Vitrification