摘要
目的:研究不同核移植方法对小鼠体细胞核移植效率的影响,建立一种简单有效的核移植方法。方法:以小鼠卵丘细胞作为体细胞核移植的供核细胞,采用4种去核方法(盲吸法、蔗糖辅助去核法、化学去核法、荧光染色去核法)研究卵母细胞去核率的差异,并比较胞质内注射法和反向核移植法应用于小鼠体细胞核移植的效果。结果:荧光染色法的去核率(88%)显著高于其他试验组(P<0.05),但囊胚率和囊胚细胞数与蔗糖辅助去核法无明显差异(P>0.05);胞质内注射法构建重构胚的效率与反向核移植法相似(P>0.05),但胞质内注射法的囊胚率和囊胚细胞数较反向核移植高(P<0.05)。结论:荧光染色去核法和胞质内注射法构建体细胞核移植胚胎效率较高,可维持胚胎早期发育。
Abstract Objective：To study the effects of different methods of nuclear transfer on the efficiency of somatic cell nuclear transfer in mouse, and establish a simple, efficient method for nuclear transfer. Methods：Cumulus cell was used as donor of mouse somatic nuclear transfer. Different enucleation methods（blind aspiration,sucrose pretreatment for enucleation, chemical induced enucleation, bisbenzimide stain method） and two methods of enucleation（intracytoplasmic nuclear injection and reverse nuclear transfer）were studied on enucleation rate of oocyte. Results. The enucleation rate（88%） was the highest in bisbenzimide stain method than in other three groups（ P〈0.05）, but there was no difference between the bisbenzimide stain and sucrose pretreatment for enucleation methods in blastocyst development and total cell numbers per blastocyst （ P 〉0.05）. The cleavage rate of reconstituted embryos between the intracytoplasmic nuclear injection and reverse nuclear transfer methods was no difference（ P〉0.05）, but intracytoplasmie nuclear injection had higher blastocyst development rate and total cell numbers per blastocyst than those of reverse nuclear transfer method（ P 〈0. 05）. Conclusion： Bisbenzimide stain method and intracytoplasmic nuclear injection are efficient procedures in mouse somatic cell nuclear transfer, these procedures have the ability to maintain early nuclear transfer embryos development.
出处
《广西医科大学学报》
CAS
2009年第2期187-190,共4页
Journal of Guangxi Medical University
基金
广西科学研究与技术开发计划课题(No.0322025-20)
广西卫生厅重点科研课题(No.重200207)
关键词
核移植
细胞质内注射
小鼠
nuclear transfer
intracytoplasmic nuclear injection
mouse
作者简介
通信作者：E-mail：mozengnan@gmail．com
