脱羰秋水仙碱诱导昆明白小鼠卵母细胞去核的研究  

Demecolcine-induced Enucleation of Kunming Mouse Oocyte

作　　者：王强[1] 安志兴[1] 顾玲[1] 马利兵[1] 郑月茂[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100

出　　处：《遗传》2004年第5期653-657,共5页Hereditas（Beijing)

基　　金：国家科技部863高技术研究发展规划(编号:2001AA213081)~~

摘　　要：在已有的demecolcine(以下简称Deme)诱导去核技术路线的基础上,以昆明白小鼠卵母细胞为实验材料,对影响去核率的几个因素(包括Deme浓度、Deme处理起始时间和作用时间、卵母细胞的卵龄)逐次进行实验。结果表明:(1)激活卵母细胞在浓度为0 4μg/mL和0 5μg/mLDeme KSOM液中处理60min均能有效去核,但0 5μg/mL组获得更高的去核率(33 3%)。(2)卵母细胞在激活后0～5min之内迅速放入0 5μg/mLDeme KSOM液中,处理60～180min得到了相对较高的去核率(31 9%～24 5%)。(3)昆明白小鼠hCG注射后17～18h收集的卵母细胞更有利于Deme诱导去核,去核率为27 1%。经比较分析,建立了优化的Deme诱导去核程序。On the basis of exiting technique pathway of demecolcine-induced enucleation(IE),several factors(Demecolcine concentration,time of demecolcine inition and treatment,oocytes age) affecting the IE rate were tested using Kunming mouse oocytes.The experiments' results demonstrated that: In experiment 1,activated oocytes could be enucleated efficiently by treating with KSOM medium containing 0.4μg/mL or 0.5μg/mL demecolcine for 60 min,but 0.5μg/mL group gained the higher IE rate(33.3%).In experiment 2,maximum IE rate (31.9%～24.5%) were obtained when oocytes were exposed to 0.5μg/mL demecolcine between 0 and 5 min postactivition and treated for 60～180 min.In experiment 3,oocytes collected from Kunming mouse at 17～18h after hCG administration were favoriate to demecolcine-IE(27.1%). By comparision and analysis of the data,we established the optimized IE procedure.

关 键 词：小鼠 诱导去核 脱羰秋水仙碱(Demecolcine) 核移植 卵母细胞 

分 类 号：Q952[生物学—动物学]