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Abstract To optimize the enucleation protocols of mouse oocytes, the study examined the effects of the enucleation methods of abradeding the zona pellucida(AZP) or sticking the zona pellucida (SZP)straightly、sucrose treatment or not、concentration of CB and inner diameter of pipettes on enucleation efficiency，and the effects of the concentration of CB on parthenogenetic embryos removed partial cytoplasm from 1/8 to 1/12. The results showed (1) there were no significant differences between supplemented 3% sucrose group and control group on manipulating time and enucleation rate achieved by AZP or SZP（P＞0.05）; (2) 5μg/mL、10μg/mL and 20μg/mL CB treatments had no significant effects on manipulating time（P＞0.05）. AZP had no significantly different enucleation rates among 5μg/mL、10μg/mL and 20μg/mL CB groups（P＞0.05）；however, in SZP, the enucleation rates of 5μg/mL、10μg/mL CB treatments were significant higher than that of 20μg/mL（P<0.05）; (3) the cleavage rate of parthenogenetic embryos had no significant difference between various groups（P＞0.05）. Blastocyst rate of 20μg/mL CB group was significant lower than that of other groups（P<0.05）; (4) the enucleation rates of 10μm、15μm and 20μm inner diameter of pipettes were no significantly different in AZP（P＞0.05）; the enucleation rates of 10μm and 15μm inner diameter of pipettes were higher than that of 20μm in SZP（P<0.05）. The enucleation rates achieved by AZP was significant higher than that achieved by SZP （P<0.05） . The results indicate that the optimized protocol combining AZP, 10μg/mL CB with 15μm inner diameter of pipettes significantly improves the efficiency of enucleation, and can be used for nuclear transfer on mouse.
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Received: 21 August 2006
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