Mouse oocyte vitrification: the effects of two methods on maturing germinal vesicle breakdown oocytes

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Abstract
Evaluation of viability and subsequent developmental ability of mouse germinal vesicle breakdown oocytes vitrified in conventional straws. Oocytes with compact cumulus cells were cultured for 3 h in TCM199 medium GVBD and vitrified by two methods: the step-wise and single-step. After vitrification, the oocytes were thawed, and subjected to in vitro maturation and in vitro fertilization. Oocyte survival (post-thaw) was assessed by morphological appearance and staining, using propidium iodide (PI)/Hoechst 33342. The oocyte maturation and fertilization rates were examined in vitro. In the single-step method the rates of post thaw survival, maturation to metaphase II and cleavage (2-cell embryos) were 58.68%, 56.41% and 38.63%, respectively. In the step-wise method, the corresponding rates were 81.75%, 68.59% and 51.80%, respectively. Vitrification of mouse germinal vesicle breakdown oocytes by the step-wise method had the advantage of maintaining the viability and subsequent production of 2-cell embryos. In comparison with that in unvitrified control oocytes, the development of MII oocytes to 2-cell embryos was impaired following vitrification.

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