小鼠脊髓前角运动神经元的分离、培养和鉴定

Isolation,culture and identification of motor neurons from embryo mouse spinal cords

目的：探讨一种分离小鼠脊髓前角运动神经元的方法,为运动神经元相关的研究提供依据.方法：分离13 d的C57BL/6胎鼠脊髓,利用Optiprep分离液经密度梯度离心获得脊髓前角运动神经元,培养后观察神经元细胞形态的变化,免疫荧光双标法对分离运动神经元的纯度进行判定.结果：将分离到的运动神经元进行培养,能够观察到典型的神经元细胞形态及轴突生长.免疫荧光双标证明培养的细胞为运动神经元细胞,纯度约95％.结论：Optiprep分离液用于小鼠运动神经元的分离,可获得良好的分选效果.Optiprep分离液可用于运动神经元相关的研究.

Objective To explore the protocol of isolation and culture of motor neurons from mouse spinal cords and provideevidence for the motor neuron-related studies. Methods： Following dissociation of E13d embryonic Ccords, motor neurons were isolated using optiprep according to the density gradient centrifugation. the cultured motor neurons were observed. Doublelabelling immunofluorescence stainning was used to identify motorneurons. Results： The cultured cell displayed typical morphology of motor neuron with neurite growth. Theimmunofluorescence assays showed that the purity of isolated motor neurons was around 95%. Conclusioeffective isolation of mouse motor neuron and can be used in the motor neuron-re