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[1]韩斌,杨芳,金锐,等.不同浓度保护剂及渗透平衡时间对慢速冷冻小鼠卵母细胞活力的影响[J].宁夏医科大学学报,2007,(01):1-3,7.
 HAN Bin,YANG Fang,WANG Yan-rong,et al.Effects of Different Concentration Cryoprotective Agents and Osmotic Equilibration Time on Biological Activation of Ultrarapid Freezing Mice Oocytes[J].Ningxia Medical University,2007,(01):1-3,7.
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不同浓度保护剂及渗透平衡时间对慢速冷冻小鼠卵母细胞活力的影响(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2007年01期
页码:
1-3,7
栏目:
论著
出版日期:
2007-01-20

文章信息/Info

Title:
Effects of Different Concentration Cryoprotective Agents and Osmotic Equilibration Time on Biological Activation of Ultrarapid Freezing Mice Oocytes
作者:
韩斌1杨芳1金锐1李灵武1王燕蓉2
1.银川市妇幼保健院生殖中心,银川 750001; 2. 宁夏医学院基础学院组织学与胚胎 学教研室,银川 750004
Author(s):
HAN Bin YANG Fang WANG Yan-rong et al
The Center for Reproductive Medicine, Women and Children Healthcare Hospital of Yinchuan City, Yinchuan 750001
关键词:
细胞保护剂 小鼠 卵母细胞 卵胞浆内单精子注射
Keywords:
cryoprotectant agent mice oocytes ICSI
分类号:
R321-33
DOI:
-
文献标志码:
A
摘要:
目的 探讨不同的冷冻保护剂浓度、渗透平衡时间对小鼠卵母细胞冷冻后存活率及 存 活卵母细胞ICSI受精后胚胎发育潜能的影响。方法 小鼠卵母细胞来自用 HMG/HCG刺激的性 成熟小鼠,依据预平衡液浓度,即1.0M 或1.5M 丙二醇(PROH),将卵母细胞分为A、B组 , 依据装载液蔗糖浓度(0.1、0.2、0.3M)的不同将A、B两组分为A1、A2、A3、 B 1、B2、B3 6组; 依卵母细胞在平衡液中的渗透平衡时间不同(<10min、>20min)分为C 、D两组; 依装 载液中的渗透平衡时间不同(<10min、>30min)分为E、F两组。卵母细胞在预平衡液中渗透 平 衡10~20min,在装载液中渗透平衡5~30min后进入慢速程序化冷冻并保存,快速解冻后观 察 卵母细胞存活率、ICSI受精率和优质胚胎形成率。结果 A、B组间冻存卵 母细胞存活率差异有非 常显著性统计学意义(43.7% vs 23.5%); A2组的卵母细胞存活率最高(52.5%),A3 组次之(42.9% ); C、D两组间冻存卵母细胞存活率、透明带破裂率差异无统计学意义。E、F两组间存活率 、透明带破损率差异有统计学意义(52.5% vs 6.3%; 6.6% vs 75.0%); 新鲜组和冷冻 组卵 母细胞受精率、卵裂率、优质胚胎率差异均有统计学意义(81.8% vs 40%; 77.3% vs 1 0 0%; 90.9% vs 83.3%)。结论(1)在慢速冷冻中,平衡液较低的PROH 浓度(1.0M)和装载液 较高的蔗糖浓度(0.2M)有益于卵母细胞冻存存活率的提高。(2)小鼠卵在平衡液中的渗透 平 衡时间以10min为宜; 在装载液中室温下的渗透平衡时间以5~10min为宜,最好不要超过15m i n。(3)冻存复苏后的存活卵经ICSI受精后受精率下降,但受精后胚胎的发育潜能似乎未受多 大影响。
Abstract:
Objective To study effects of different cryoprotective solutio n conce ntration and osmotic equilibration time on the survival rate of frozen-thawed m i ce oocytes and the developmental potential of the embryos that the survival oocy tes were fertilized by ICSI. Methods Mice oocytes were obtained from the mice th at were treated by ovarian stimulation protocol with HMG/HCG. By equilibration s olution concentration, oocytes were divided into A(1.0M PROH)and B(1.5M PROH) g roup. Then by loading solutions( sucrose)of different concentration(0.1M, 0 .2 M, 0.3M), A group and B group were respectively divided into A1,A2, A3 group a n d B1,B2,B3 group. By different osmotic equilibration time of equilibration solu t ions, oocytes were divided into C(<10min)and D(>20min)group. By different o smo tic equilibration time of loading solutions, oocytes were divided into E(<10min ) and F(>30min)group. Mice oocytes were slowly frozen by an automated biologic al freezer and stored in liquid nitrogen tanks after the mice oocytes were osmotic ally equilibrated within 10~20min in the equilibration solutions and within 5~ 30 min in the loading solutions. The survival rate,fertilization rate and high qu a lity embryos rate of oocytes post-thawed rapidly by ICSI were observed. Results The survival rate of frozen oocytes in A group was significantl y higher than th at in B group. No difference of survival rate of frozen oocytes and broken rate of zona between C and D group was found. The survival rate of frozen oocytes in E group was significantly higher than that in F group and the broken rate of zon a in E group was significantly lower than that in F group. The fertilization ra t e, cleavage mice and high quality embryo mice in fresh group were significantly higher than that in frozen group. Conclusion(1)As slow frozen, lower concentrat ion PROH(1.0M)of the equilibration solution and higher concentration sucrose ( 0.2M)of the loading solution were beneficial for increasing the survival rate o f cryopreservation mouse oocytes.(2)The osmotic equilibration time of oocytes i n the equilibration solution was suitable in 10 minutes, and its time in the loa ding solution in the room temperature was also suitable in 5-10 minutes and had better not to exceed 15 minutes.(3)Although the fertilization rates of oocytes p ost-thawed by ICSI was low, the developmental potential of embryos seemed not to be influenced.

参考文献/References:

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[11] 李梅,陈子江.慢速冷冻对人成熟卵与未成熟卵发育潜能的影响[J].实 用妇产科杂志,2003,19(3):147-149.

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备注/Memo

备注/Memo:
收稿日期:2006-06-21 基金项目:银川市科技局资助(2004-34) 作者简介: 韩斌(1962-),男,宁夏人,副主任医师,硕士,从事生殖医学研究。 通讯作者:王燕蓉,教授,硕士研究生导师,从事低温生物学研究。E-mail:4083304 @163.com
更新日期/Last Update: 2007-01-20