原代小胶质细胞及神经元细胞培养不同纯化方法的比较

Comparison of different purified methods in culture of primary microglia and neuron culture

目的探讨建立简便高效的原代小胶质细胞及神经元培养和纯化方法。方法神经元与胶质细胞混合培养后,采用温和胰酶消化法、恒温振荡法、利多卡因分离法及手摇法分离纯化小胶质细胞,无血清培养法及阿糖胞苷法分离纯化神经元,细胞计数、流式细胞术及细胞化学染色鉴定小胶质细胞及神经元。结果手摇法分离纯化小胶质细胞与温和胰酶消化法、恒温振荡法、利多卡因分离法比较,细胞产量更高、活性更好;无血清培养法培养神经元比阿糖胞苷法细胞纯度更高、活性更好,形成神经网络结构的时间也更早。结论手摇法分离纯化小胶质细胞是产量最高、简便、经济的方法,而无血清培养法更能提高神经元的纯度及活性。

Objective To establish an easy and effective method for cuhure and purification of primary microglia and neurons. Methods Mild trypsinization, mechanical shaking （37℃ 2 h at 250 r/min）, lidocaine hydrochloride and hand shaking methods were applied for isolation and purification of glias from mixed glias and neurons culture, while neu- rons with neurobasal medium and cytarabine dealing methods. All the purified cells were counted. The purity of isolated microglia and neuron were assessed by flow cytometriy and immunocytochemitry, respectively. Results The hand shaking method for isolation of microglias was proved with highest efficacy and vivacity, and so was the neurobasal medium method for neurons. Conclusion It＇s a productive and economic method of hand shaking method in purification of microglia, and so is neurobasal culture method for neurons.