脱羰秋水仙碱诱导昆明白小鼠卵母 细胞去核的研究

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.

Demecolcine-induced Enucleation of Kunming Mouse Oocyte

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 0.4 microg/mL or 0.5 microg/mL demecolcine for 60 min, but 0.5 microg/mL group gained the higher IE rate(33.3%).In experiment 2, maximum IE rate (31.9% -approximately 24.5%) were obtained when oocytes were exposed to 0.5 microg/mL demecolcine between 0 and 5 min postactivation and treated for 60 -approximately 180 min.In experiment 3,oocytes collected from Kunming mouse at 17 -approximately 18h after hCG administration were favoriate to demecolcine-IE(27.1%). By comparison and analysis of the data, we established the optimized IE procedure.