大鼠脊髓背根神经节神经元的纯化培养

Purification of dorsal root ganglion neurons in rat

目的:建立一种简单、稳定、高效的大鼠脊髓背根神经节神经元原代纯化培养方法。方法:取新生SD大鼠的脊髓背根神经节,采用胰蛋白酶消化法,制成单细胞悬液,加入阿糖胞苷以纯化细胞,培养于含10%胎牛血清与重组人胶质细胞源性神经营养因子的DF12培养基中,观察神经元生长状况,用细胞计数和神经元特异性烯醇化酶(NSE)免疫细胞化学染色测定细胞纯度。结果:培养的脊髓背根神经节神经元生长状态正常,有神经突起生长和标记蛋白的表达,可达到90%左右的纯度。结论:本培养方法简单、稳定、高效,为对神经元进一步的深入研究提供了实验模型。

Objective：To establish an easy,practical,reliable method for the purification culture system of dorsal root ganglion neurons derived from rats.Method：Dorsal root ganglions harvested from newly-born rats were digested with trypsin and produced into single cell suspension,then plated in DF12 media containing 10% FBS and glial cell line-derived neurotrophic factor （GDNF）.Cultured dorsal root ganglion neurons were purified by cytosine arabinoside for 48 hours.The purificational rates were evaluated according to cell count and neuronal specific enolase （NSE） immunocytochemistry stain,Result：Cultured dorsal root ganglion cells could survive healthily,The purification rate of neurons was 90% or so,Conclusion：The dorsal root ganglion neurons cultivated in the DF12 media with fetal bovine serum and GDNF can survive with nomal cell phenotype,neurite synapse growth and the expression of neurifilament protein is available.Which is a useful model for the further studies,