小鼠卵母细胞玻璃化冷冻：两种方法对生发泡期卵母细胞成熟的影响。
Mouse oocyte vitrification: the effects of two methods on maturing germinal vesicle breakdown oocytes.
机构信息
Department of Anat., Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
J Assist Reprod Genet. 2010 May;27(5):233-8. doi: 10.1007/s10815-010-9411-x. Epub 2010 Apr 21.
PURPOSE
Evaluation of viability and subsequent developmental ability of mouse germinal vesicle breakdown oocytes vitrified in conventional straws.
METHODS
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.
RESULTS
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.
CONCLUSION
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.
目的
评估传统 straw 中玻璃化的小鼠生发泡破裂卵母细胞的活力和随后的发育能力。
方法
用 TCM199 培养基培养紧密的卵丘细胞 3 小时，使其生发泡破裂，采用逐步法和一步法进行玻璃化处理。玻璃化处理后，将卵母细胞解冻，进行体外成熟和体外受精。通过碘化丙啶（PI）/Hoechst 33342 染色，根据形态外观评估卵母细胞的存活（解冻后）。体外检查卵母细胞成熟和受精率。
结果
在一步法中，解冻后存活率、中期 II 成熟率和卵裂（2 细胞胚胎）率分别为 58.68%、56.41%和 38.63%。在逐步法中，相应的比率分别为 81.75%、68.59%和 51.80%。
结论
用逐步法对小鼠生发泡破裂卵母细胞进行玻璃化处理有利于维持其活力和随后产生 2 细胞胚胎。与未经玻璃化处理的对照组卵母细胞相比，玻璃化处理后，MII 期卵母细胞发育为 2 细胞胚胎的能力受损。
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