[1]海兰）,谭丽）#,毛跟红）,等.不同冷冻载体对小鼠成熟卵母细胞玻璃化冷冻的影响[J].郑州大学学报(医学版),2012,(04):550.
　HAI Lan,TAN Li,MAO Genhong,et al.Effect of different cryopreservative carriers on vitrification of mouse MⅡ oocytes[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(04):550.
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不同冷冻载体对小鼠成熟卵母细胞玻璃化冷冻的影响
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2012年04期
页码:
550
栏目:
应用研究
出版日期:
2012-07-20

文章信息/Info

Title:
Effect of different cryopreservative carriers on vitrification of mouse MⅡ oocytes
作者:
海兰1）; 谭丽2）#; 毛跟红2）; 马丽影2）; 赵冬梅2）; 项云改2）; 李艳2）
1）美国南伊利诺伊大学医学院生理系 卡本戴尔 62901 2）郑州大学第二附属医院生殖中心 郑州 450014
Author(s):
HAI Lan1）; TAN Li2）; MAO Genhong2）;  MA Liying2）; ZHAO Dongmei2）; XIANG Yungai2）; LI Yan2）
1) Department of Physiology,School of Medicine，Southern Illinois University, Carbondale 62901 2) Reproductive Medical Center, the Second Affiliated Hospital,Zhengzhou University,Zhengzhou 450014
关键词:
玻璃化冷冻; 冷冻载体; 普通麦管; 拉细麦管; 卵母细胞; 体外受精
Keywords:
vitrification; cryopreservative carrier; regular straw; pulled straw; oocyte; in vitro fertilization
分类号:
R339.2
摘要:
目的：探讨玻璃化冷冻小鼠成熟减数第二次分裂中期（M Ⅱ期）卵母细胞的最佳冷冻载体及冷冻方法。方法：用乙二醇和蔗糖为玻璃化冷冻保护剂，将小鼠成熟M Ⅱ期卵母细胞分为冻融、暴露和对照组。冻融组分别采用普通麦管、封闭式拉细麦管（CPS）和开放式拉细麦管（OPS）作载体，采用玻璃化快速冷冻和快速复温的方法冻存小鼠成熟卵母细胞；暴露组卵母细胞仅暴露在冷冻和复苏液中，不进行冷冻；对照组卵母细胞不接触冷冻和复苏液。观察各组处理后卵子回收、受精、卵裂等情况。结果：冻融组普通麦管、CPS和OPS处理的卵母细胞回收率分别为92.0%、92.3%和72.3%，OPS处理卵母细胞的回收率显著低于普通麦管和CPS处理的卵母细胞（P<0.001）；普通麦管、CPS和OPS处理的卵母细胞复苏存活率分别为43.5%、72.2%和76.5%，CPS和OPS处理卵母细胞的复苏存活率显著高于普通麦管组处理的卵母细胞（P<0.001）。冻融、暴露和对照组的卵裂率分别为38.6%、69.2%和73.5%，冻融组显著低于暴露和对照组（P<0.001）。结论：CPS法在玻璃化冷冻保存成熟的卵母细胞方面有价值。
Abstract:
Aim:To explore the optimal carrier and procedure for mouse mature (MⅡ) oocyte vitrification.Methods:Collected mouse MⅡ oocytes and randomly divided into experimental,exposed,and control groups.Three types of carriers［regular straws,closed pulled straws (CPS),and open pulled straws (OPS)］ were used in expreimental group.Ethylene glycol and sucrose were used as cryopreservatives to flash freeze and thaw the oocytes in experimental group. The oocytes in the exposed group were only exposed to ethylene glycol and sucrose but not frozen and the oocytes in the control group were not treated.The postthawed oocyte retrieval rate, fertilized rate, and cleavage rate were observed after vitrification.Results:Oocyte retrieval rates were 92.0%, 92.3%, and 72.3% in regular straws, OPS, and CPS treatment oocytes, respectively. Oocyte retrieval rate in the CPS treatment oocytes was much lower compared with the other two treatment oocytes (P<0.001). Postthawed oocyte survival rates were 43.5%, 72.2%, and 76.5% in regular straws,CPS,and OPS treatment oocytes,respectively.The rates of CPS and OPS treatment oocytes were much higher in comparison to the regular straw treatment oocytes(P<0.001). The cleavage rates in experimental, exposed, and control groups were 38.6%, 69.2%, and 73.5% ,respectively. The cleavage rate of experimental group was much lower than those of the exposed and control groups (P<0.001).Conclusion: CPS is a promising carrier in vitrification for mouse MⅡ oocytes and worth trying to put into application.

参考文献/References:

［1］Zhou XL,Al Naib A,Sun DW,et al. Bovine oocyte vitrification using the cryotop method: effect of cumulus cells and vitrification protocol on survival and subsequent development［J］.Cryobiology，2010，61(1):66
[2]罗丽兰.不孕与不育［M］.2版.北京：人民卫生出版社,2009.
[3]Stachowiak EM, Papis K, Kruszewski M,et al.Comparison of the level(s) of DNA damage using comet assay in bovine oocytes subjected to selected vitrification methods［J］. Reprod Domest Anim,2009,44(4):653
[4]卢惠霖, 卢光琇.人类生殖与生殖工程［M］.郑州：河南科学技术出版社,2001.
[5]严正杰，蔡令波，冯婷，等.卵母细胞玻璃化冷冻对胚胎发育及妊娠结局的影响［J］.生殖医学杂志,2009,18(1):17
[6]Liu L,Milroy C,Peterson CM,et al.Successful cryoloop vitrification and subsequent in vitro maturation of mouse preantral follicles［J］. Syst Biol Reprod Med,2011,57(3):149
[7]Smith GD, Serafini PC, Fioravanti J,et al.Prospective randomized comparison of human oocyte cryopreservation with slowrate freezing or vitrification［J］. Fertil Steril,2010,94(6):2088
[8]Chen SU,Lien YR,Chen HF,et al.Vitrification of mouse oocytes using CPS achieves a high survival and preserves good patterns of meiotic spindles, compared with conventional straws, OPS and grids［J］. Hum Reprod,2001,16(11):2350
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备注/Memo

备注/Memo:
#通讯作者，女，1964年3月生,博士,教授,研究方向:生殖内分泌,Email:litan668@zzu.edu.cn
更新日期/Last Update: 2012-08-14