遗传 ›› 2004, Vol. 26 ›› Issue (5): 653-657.

• 论文 • 上一篇    下一篇

脱羰秋水仙碱诱导昆明白小鼠卵母 细胞去核的研究

王 强; 安志兴; 顾 玲; 马利兵; 郑月茂; 张涌WANG Qiang; AN Zhi-Xing ; GU Ling ; MA Li-Bing ; ZHENG Yue-Mao ;ZHANG Yong   

  1. 西北农林科技大学生物工程研究所,陕西杨凌 712100Institute of Bio-Engineering Northwest Sci-Tech University of Agriculture and Forestry,Shaanxi Yangling 712100,China
  • 收稿日期:1900-01-01 出版日期:2004-10-10 发布日期:2004-10-10

Demecolcine-induced Enucleation of Kunming Mouse Oocyte

  • Received:1900-01-01 Online:2004-10-10 Published:2004-10-10

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/mL Deme-KSOM液中处理60 min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0~5 min之内迅速放入0.5μg/mL Deme-KSOM液中,处理60~180 min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17~18 h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。

Abstract: 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: 脱羰秋水仙碱(Demecolcine), 核移植, 卵母细胞 Key word, 诱导去核, 小鼠