Gao Shanshan, Li Yuan, Gao Xuan, Hu Jingmei, Yang Huijun, Chen Zi-Jiang
Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, China.
Reprod Sci. 2009 Apr;16(4):391-6. doi: 10.1177/1933719108327590. Epub 2008 Dec 15.
This experiment investigated the optimal time required for cryopreserved human oocytes to reform their spindles upon re-warming. Metaphase II oocytes were cryopreserved with a slow freezing method. Oocytes from each patient were randomly allocated into one of the 3 groups with different culture periods: 1, 3, or 5 hours, respectively, after thawing. Tubulin and chromosome configurations were visualized by confocal microscopy after immunostainings. By morphological assessment, 87.3% oocytes survived the process of freezing and thawing. Oocytes with normal spindle configuration increased significantly after 3 or 5 hours of incubation compared to those incubated for only 1 hour ( P < 0.05). There were no differences in the chromosome configurations among the treatment groups ( P > 0.05). This experiment demonstrated that cryopreserved human oocytes need a certain minimum period of incubation time (3 h) to recover their disrupted MII spindles and this information can be used in development of human IVF protocols with frozen oocytes.
本实验研究了冷冻保存的人类卵母细胞复温后重新形成纺锤体所需的最佳时间。采用慢速冷冻法对处于减数分裂中期II的卵母细胞进行冷冻保存。解冻后，将每位患者的卵母细胞随机分配到3组中，分别培养1、3或5小时。免疫染色后，通过共聚焦显微镜观察微管蛋白和染色体的形态。通过形态学评估，87.3%的卵母细胞在冻融过程中存活下来。与仅培养1小时的卵母细胞相比，培养3或5小时后，纺锤体形态正常的卵母细胞显著增加（P<0.05）。各处理组之间的染色体形态没有差异（P>0.05）。本实验表明，冷冻保存的人类卵母细胞需要一定的最短培养时间（3小时）来恢复其受损的减数分裂中期II纺锤体，该信息可用于制定冷冻卵母细胞的人类体外受精方案。
