一种改进的高密度大鼠皮层神经元培养方法  

An improved method for culture of high-density rat cortex neurons

作　　者：康国创[1] 刘文博[1] 尹丽鹤[2] 费舟[1] 

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032 [2]第四军医大学西京医院神经内科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2009年第5期417-420,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30670796)

摘　　要：目的建立一种便于量化操作且纯度较高的大鼠皮层神经元培养方法,并观察其生长的形态学变化规律。方法采用精确控制胎龄的SD大鼠胚胎,在低温磷酸盐缓冲液(PBS)中彻底剥离脑膜组织后取双侧皮层,充分剪碎消化,以600～700细胞/mm2的密度接种于预处理过的固相培养载体上,并对不同培养时间的神经元进行形态学观察和纯度分析。结果利用本方法成功培养出含杂质量较少且纯度很高的高密度皮层神经元,其在不同生长阶段具有典型的形态学特征。结论该方法可以顺利的培养出纯度很高的高密度生长的皮层神经元,便于量化控制操作步骤,减少操作误差,为今后相关研究奠定了方法学基础。Objective To establish an improved method for culture of high-density fetal cortex neuron and observe morphological characteristics of the neurons at different developmental and differential stages. Methods The cortex of precise 14 d embryonic rat was used for culture in this study. Shortened time of trypsin digestion and mechanical dissociation were adopted to reduce the damage of neurons during the operation. Then the single-neuron-cell suspension was planted to poly-L-lysin coated plate with a density of 600-700 cells/mm^2 and the neuronal morphological characteristics at different stages were observed. Results High-density rat cortex neurons were successfully cultured, which had distinct morphological characteristics at different stages. Conclusion This high-density culture method is a stable method for neurons in vitro research.

关 键 词：高密度 皮层 神经元 培养 方法 

分 类 号：R446.6[医药卫生—诊断学]