建立一种简便的大鼠背根神经节神经元原代培养方法的建立

发布时间：2018-04-05 21:29

  本文选题：背根神经节(DRG)　切入点：原代培养　出处：《神经解剖学杂志》2014年02期

【摘要】：目的:建立一种简便易行地获取大鼠背根神经节(dorsal root ganglion,DRG)神经元原代培养的方法。方法:窒息法处死大鼠,打开胸腔经心脏放血后,从背部取出一段脊柱。将脊柱稍加修剪后,从中轴线处打开脊柱,暴露脊髓,在其两侧找到椎间孔内的为DRG,用器械离断神经纤维,取出DRG。Ⅰ胶原酶联合Trypsin-EDTA先消化DRG,再用吹法DRG以获得单个神经元;用TNB-100 basal Medium的复合培养液在经过0.1 mg/ml PolyD-lysine hydrobromide和0.01 mg/ml lamina包被的培养皿内,于5%CO2,37℃孵箱内培养48 h。免疫组织化学方法检测神经元内标志物BetaⅢTubulin的表达,用Alexa flour 488标记的二抗进行显色,倒置荧光显微镜下观察。结果:获得了单层贴壁的神经元,其胞体形状多样,突起生长良好,细胞之间正在逐步形成网络连接;免疫荧光染色显示,细胞的胞体和突起中均可高表达BetaⅢTubulin,且荧光染色较强。结论:运用该方法可以简便地从大鼠DRG内获得单层贴壁的原代神经元。
[Abstract]:Aim: to establish a simple and convenient method for the primary culture of dorsal root ganglion (DRG) neurons in rat dorsal root ganglia (DRG).Methods: the rats were killed by asphyxia.After a little pruning of the spine, the spinal column was opened from the central axis and the spinal cord was exposed. The nerve fibers in the intervertebral foramen were found on both sides of the spinal column. The nerve fibers were cut off, and the DRG鈪，

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