本文選題：背根神經(jīng)節(jié)(DRG)　切入點(diǎn)：原代培養(yǎng)　出處：《神經(jīng)解剖學(xué)雜志》2014年02期

【摘要】：目的:建立一種簡(jiǎn)便易行地獲取大鼠背根神經(jīng)節(jié)(dorsal root ganglion,DRG)神經(jīng)元原代培養(yǎng)的方法。方法:窒息法處死大鼠,打開胸腔經(jīng)心臟放血后,從背部取出一段脊柱。將脊柱稍加修剪后,從中軸線處打開脊柱,暴露脊髓,在其兩側(cè)找到椎間孔內(nèi)的為DRG,用器械離斷神經(jīng)纖維,取出DRG。Ⅰ膠原酶聯(lián)合Trypsin-EDTA先消化DRG,再用吹法DRG以獲得單個(gè)神經(jīng)元;用TNB-100 basal Medium的復(fù)合培養(yǎng)液在經(jīng)過0.1 mg/ml PolyD-lysine hydrobromide和0.01 mg/ml lamina包被的培養(yǎng)皿內(nèi),于5%CO2,37℃孵箱內(nèi)培養(yǎng)48 h。免疫組織化學(xué)方法檢測(cè)神經(jīng)元內(nèi)標(biāo)志物BetaⅢTubulin的表達(dá),用Alexa flour 488標(biāo)記的二抗進(jìn)行顯色,倒置熒光顯微鏡下觀察。結(jié)果:獲得了單層貼壁的神經(jīng)元,其胞體形狀多樣,突起生長(zhǎng)良好,細(xì)胞之間正在逐步形成網(wǎng)絡(luò)連接;免疫熒光染色顯示,細(xì)胞的胞體和突起中均可高表達(dá)BetaⅢTubulin,且熒光染色較強(qiáng)。結(jié)論:運(yùn)用該方法可以簡(jiǎn)便地從大鼠DRG內(nèi)獲得單層貼壁的原代神經(jīng)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，

本文編號(hào)：1716558