摘要
目的:建立一种简单高效的中脑多巴胺神经元细胞原代培养方法,并观察胰酶消化对中脑多巴胺能神经元突起生长的损伤作用。方法:以Nakai等经典神经元细胞原代培养方法为基础,通过使用低日龄胎鼠,初次培养液加入胎牛血清等步骤,促进中脑多巴胺能神经元细胞贴壁和生长;在无胰酶消化组直接使用内口外翻的小口径硅化吸管轻柔吹打离散细胞,比较两种方法间神经元细胞突起形成的差异。结果:接吹打组其多巴胺能神经元细胞突起的生长程度(212±10μm)明显高于胰酶消化组(113±9μm)(P<0．01),而两组间多巴胺阳性细胞比例未见显著差异(P>0．05)。结论:在中脑多巴胺能神经元细胞原代培养中,低日龄胎鼠及免胰酶消化离散细胞可减少神经元细胞损伤,有利于细胞突起的生长。
Objective： To establish an easy and hlgh-field method of rat dopaminergic neuron primary culture, and to observe the damage to the neurites by the trypsogen. Methods： Based on classic Berger＇s primary cell culture method, We used low conceptus age rat embryos, and add the fetal bovine serum into the culture solution for the first time, to increase the dopaminergic neuron adherence and growth; In the group of direct separation, we use small caliber siliclfication tube which＇s canal orifice was everted to blow the tissue.and compare the difference about neurids between two culture methods. Results： In the group of direct blow, the dopaminergic neuron neurifis＇s length （212±10μm） is longer than the group of trypsinizadon （113±9μm） （P〈0.01） significandy. And the percentage of TH positive neurons between two groups is no significant deviation （P〉0.05）. Conclusion： The low conceptus age rat embryos and direct separation can decrease the damage to the primary cell culture and make the cell neurids well stretching.
出处
《现代生物医学进展》
CAS
2006年第12期10-12,共3页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30571545)
