Successful In Vitro Maturation of Oocytes Collected from Whole Porcine Ovaries Vitrification

摘要

Background: Ovarian cryopreservation has become more and more important not only as a tool of genetic resource and biodiversity conservation but also as a bio-medical application. Objective: The aim of the present study is to survey on slow freezing and vitrification method towards a simple method to cryopreserve intact ovary which can be applied in the field condition. The result of method base on evaluation of viability and maturation potential of immature oocytes were collective from frozen–thawed ovaries. Materials and methods: The whole porcine ovarian were vitrified using same cryoprotective solution in three groups (V1, V2 and V3) which the treatment time in ES solution is 15, 30 and 45 min, respectively comparing to slow freezing program. After three weeks of storage, frozen samples were thawed and immature oocytes were collected for in vitro maturation. FDA staining of immature oocytes was used to con?ˉ????rmed the viability and morphology of intracellular structures. The oocytes matured were evaluated by extrusion of the first polar body. Results: The percentage of viability oocytes was the highest in group V2 (7.03%) compare with that in other group V1; control group and V3 group (6.47; 0.91 and 0.65%, respectively). The maturation rate did differ in two groups V1 (6.47%) and V2 (7.03%), but it was not in control and V3. Conclusion: whole ovarian vitrified is already a feasible technique, it is possible to collect viable immature oocytes that have the ability to mature in vitro.