目的：通过检测玻璃化冷冻前后卵母细胞线粒体DNA（mtDNA）缺失和能量代谢变化，以探讨冷冻保存对卵母细胞质量的影响。方法：收集体外受精（IVF）废弃的人卵母细胞和受精卵，包括未受精（0PN）、两原核（2PN）和三原核（3PN）；随机分为2组，A组为经玻璃化冷冻保存，B组为新鲜对照组。采用巢式聚合酶链反应（nest PCR）检测冷冻前后卵母细胞mtDNA缺失差异；并进一步比较冷冻前后卵母细胞mtDNA拷贝数和三磷酸腺苷（ATP）含量变化。结果：卵母细胞mtDNA缺失率和mtDNA拷贝数在冷冻处理前后差异无统计学意义（P＞0.05）；冷冻后卵母细胞内ATP含量低于冷冻前新鲜卵母细胞。结论：冷冻保存对卵母细胞mtDNA缺失及拷贝数影响较小，可较好地实现保存卵母细胞的目的。%Objective: To explore the effect of cryopreservation on the mitochondrial DNA (mtDNA) deletions and energy metabolism of human oocyte, so as to investigate the effect of cryopreservation on oocytes preservation. Methods: The dumped oocytes with 0PN, 2PN and 3PN during routine IVF were collected for this cryopreservation study. Those oocytes were randomly divided into two groups, one for vitrification cryopreservation (Group A) and the other as control (Group B, without cryopreservation). The nested polymerase chain reaction (PCR) was used to detect the mtDNA deletions of oocyte mtDNA before and after cryopreservation. Real-time quantitative PCR was adopted to determine the copy number of oocyte mtDNA. ATP contents were detected using luciferin-luciferase assays. Results: There were not significant differences in the mutation frequency of oocyte mtDNA deletions and the copy number of mtDNA between two groups (P>0.05). However, The ATP content in the group A was significantly lower than that in the group B. Conclusions: The effect of cryopreservation on the oocyte mtDNA deletions and the copy number of mtDNA was limited, suggesting that cryopreservation can be properly used to preserve oocyte.
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