目的：建立一种简单、稳定、高效的新生大鼠背根神经节神经元原代培养方法。方法：摘取新生24h SD大鼠背根神经节，采用0.25%胰酶和0.1%Ⅳ型胶原酶消化，制成单细胞悬液，接种于Neurobasal/B27无血清培养液中。将培养3d的DRGn于倒置相差显微镜下进行形态学观察，扫描电镜行细胞形态学检测，应用β-tubulinⅢ进行免疫细胞化学染色，鉴定细胞纯度。结果：体外培养的背根神经节神经元生长状态良好，纯度可达到(92±6)%。结论：本实验方法简单、稳定、高效，可以获得高纯度的背根神经节神经元。%Objective To establish an simple, efficient, reliable method for the purification culture system of dorsal root ganglion neurons derived from new born rats.Methods Dorsal root ganglions harvested from new born SD rats were digested with the mixture of trypsin and collegoneaseⅣ, then turned into single cell suspension and plated in neuralbasal media. The purified rate was evaluated according to cell count and β-tubulinⅢ immunocytochemistry stain.Results Cultured dorsal root ganglion cells could survive healthily. The purification rate of neurons was(92±6)%.Conclusion The method, which is used for culture and purification of DRGn, is a simple, efficient and reliable way. Using it could obtain highly purify neurons.
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