[两种不同玻璃化冷冻保存方法对小鼠卵母细胞纺锤体的影响]

[Influence of two different vitrification cryopreservation methods on spindles of mouse oocytes].

作者信息

Sun Hai-xiang, Hu Ya-li, Chen Lin-jun, Zhang Ning-yuan, Xu Zhi-peng

机构信息

Reproductive Medicine Center, Drum Tower Hospital, Nanjing University Medical College, Nanjing, Jiangsu 210008, China.

出版信息

Zhonghua Nan Ke Xue. 2006 Dec;12(12):1076-9, 1083.

PMID:17201250

Abstract

OBJECTIVE

To investigate the influence of two different vitrification cryopreservation methods on the spindles of mouse M II oocytes.

METHODS

Three groups were included in the experiment, Group A, Group B and the control ( fresh oocytes). Mouse oocytes were vitrified by using cryoloop, with ethylene glycol( EG) in Group A and with EG + dimethyl sulphoxide ( DMSO) in Group B as cryoprotectants, and then the oocytes were placed directly into liquid nitrogen. Three hours after the frozen oocytes were thawed they were fixed, and the microtubule and chromosome were stained by indirect immunofluorescent method.

RESULTS

The survival rates of the oocytes after treated by the two vitrification cryopreservation methods had no difference ( 80. 3% vs 87. 5% , P > 0. 05) . The rate of the intact spindles in Group A was much lower than that of the control and Group B ( 15. 2% vs 78.7% , 15. 2% vs 77. 5% , P < 0. 05). But there was no difference between the latter two groups (78. 7% vs 77. 5% , P >0. 05). The oocytes with normal chromosome in Group A were much less than in the control and Group B (17.4% vs 76. 6% , 17. 4% vs 72. 5% , P <0. 05) , with no difference between the latter two groups(76. 6% vs 72. 5% , P >0. 05) ; The oocytes with abnormal chromosome were more in Group A than in the control and Group B (82. 6% vs 19. 1% , 82. 6% vs 27. 5% , P <0. 05) , with no difference between the latter two groups (19.1% vs 27.5% , P >0.05).

CONCLUSION

The changed vitrification cryopreservation method helps conserve the intact spindle configuration of mouse oocytes.

摘要

目的

探讨两种不同的玻璃化冷冻保存方法对小鼠MⅡ期卵母细胞纺锤体的影响。

方法

实验分为A组、B组和对照组（新鲜卵母细胞）。采用冷冻环法对小鼠卵母细胞进行玻璃化冷冻，A组以乙二醇（EG）作为冷冻保护剂，B组以乙二醇+二甲基亚砜（DMSO）作为冷冻保护剂，然后将卵母细胞直接放入液氮中。冷冻卵母细胞解冻3小时后进行固定，采用间接免疫荧光法对微管和染色体进行染色。

结果

两种玻璃化冷冻保存方法处理后的卵母细胞存活率无差异（80.3%对87.5%，P>0.05）。A组完整纺锤体的比例远低于对照组和B组（15.2%对78.7%，15.2%对77.5%，P<0.05）。但后两组之间无差异（78.7%对77.5%，P>0.05）。A组染色体正常的卵母细胞远少于对照组和B组（17.4%对76.6%，17.4%对72.5%，P<0.05），后两组之间无差异（76.6%对72.5%，P>0.05）；A组染色体异常的卵母细胞多于对照组和B组（82.6%对19.1%，82.6%对27.5%，P<0.05），后两组之间无差异（19.1%对27.5%，P>0.05）。