[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.