小鼠背根神经节神经细胞的原代培养新方法
A new method for primary culture of mouse dorsal root ganglion neurons

摘要：目的 建立一种操作简便、高纯度的小鼠背根神经节神经细胞原代培养的方案。方法 取6～8周龄健康雄性C57BL/6小鼠的背根神经节，使用Ⅰ型胶原酶和胰蛋白酶消化，获取背根神经节神经细胞，采用小鼠神经元特异性烯醇化酶(NSE)单克隆抗体免疫细胞化学染色鉴定并测定细胞纯度。结果 培养的原代神经细胞生长良好，纯度可达90%，用含有神经生长因子(NGF)的DMEM培养基培养，存活时间可达60d。结论 该方案简单稳定，可培养出大量高纯度的神经细胞，为深入研究神经细胞提供了可靠的模型。
ABSTRACT: Objective To establish a simple method for primary culture of mouse dorsal root ganglion neurons of high purity. Methods The dorsal root ganglions from healthy C57BL/6 mice of 6-8 weeks were taken to obtain dorsal root ganglion neurons by using type Ⅰ collagenase and trypsin digestion. Identification and purification were evaluated by neuron specific enolase (NSE) monoclonal antibody immunocytochemistry staining. Results The cultured primary neurons grew well and the purity could reach about 90%. The survival time was 60 days when cultured with the DMEM medium containing nerve growth factor (NGF). Conclusion The culture program is simple and stable, and can cultivate a large number of high-purity neurons, which provides a reliable model for in-depth study of neurons