小鼠耳蜗螺旋神经元的原代培养及鉴定  

Primary Culture in Vitro and Identification of Murine Spiral Ganglion Neurons

作　　者：刘涛[1] 陈艳芳[2] 刘义 唐玥玓[1] 

机构地区：[1]四川大学华西医院耳鼻咽喉科,四川成都610041 [2]四川大学华西医院麻醉与危重急救研究实验室,四川成都610041 [3]广东省云浮市人民医院耳鼻喉科,广东云浮527300

出　　处：《华西医学》2008年第1期104-105,共2页West China Medical Journal

基　　金：国家自然科学基金资助项目(编号30471880)

摘　　要：目的:机械分离、培养小鼠耳蜗螺旋神经元,并进行免疫荧光细胞学鉴定,为后期进一步的实验研究提供实验材料。方法:采用初出生1～5天以内的昆明小鼠进行解剖、机械分离以获得螺旋神经节组织,进行原代培养后,应用神经微丝蛋白(Neurofilament protein,NFP-H)单克隆抗体进行免疫荧光细胞学鉴定。结果:机械分离后获得的螺旋神经节组织中的螺旋神经元,在体外培养条件下可以存活并进行正常分化。典型的螺旋神经元,其细胞形态呈椭圆形,胞体透明光滑、接近生理形态。荧光染色标记后,胞体和神经突起均显色好,Schwann细胞和成纤维细胞未着色。结论:应用机械分离的方法获得小鼠耳蜗螺旋神经节组织并进行培养,耳蜗螺旋神经元在体外可以稳定地存活生长。培养获得的细胞形态和生存状态接近生理状态,满足电生理、免疫细胞化学、药理学等研究。应用特异性的神经微丝蛋白对培养获得的螺旋神经元进行免疫荧光细胞学鉴定,特异性好,荧光显色好。Objective： Information on the culture model of spiral ganglion neurons of murine cochlear was presented. Method： Cochlear of Kunming mouse which born short of five days were dissected and separated to acquire the tissue of spiral ganglion which were passed primary culture and identified by neurofilament protein NFP- H monoclonal antibody. Resnlt： Ideal spiral ganglion neurons were acquired by primary culture in vitro without enzyme. The typical cell shape of the acquired spiral ganglion neurons was ellipse and the cell body was transparent and slick. Conclusion： The acquired spiral ganglion neurons without enzyme can be primary culture and can be identified by immunocytochemical method and suit cytological experiments.

关 键 词：螺旋神经元 原代培养 神经微丝蛋白 免疫荧光细胞学 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]