机械分离并培养螺旋神经节神经元方法的建立

目的　建立机械分离并培养小鼠耳蜗螺旋神经节神经元 (SGN )细胞模型的方法。方法　对出生后1～ 6 d的小鼠耳蜗螺旋神经节组织进行机械分离和培养 ,用倒置相差显微镜对原代培养的 SGN细胞的形态学和轴突生长情况进行观察 ,并应用荧光显微镜对 SGN进行免疫细胞化学染色鉴定。结果　新生小鼠 SGN在机械分离、体外培养条件下 ,可以获得良好的细胞形态和轴突再生能力 ,其存活细胞的数量为 (78.10± 4 .33) / cm2 (5～ 7d) ,生长周期为 7～ 14 d。结论　成功建立了机械分离并培养 SGN细胞模型的方法 ,SGN存活细胞的数量和周期可以满足体外多种实验的需要。

Establishment of Methods of Mechanical Dissociation and Culture of Spiral Ganglion Neurons in Postnatal Mice

OBJECTIVE: To conduct a study on the methods for establishing a cytological model of mechanical dissociation and culture of spiral ganglion neurons (SGNs) in postnatal mice. METHODS: The spiral ganglion neurons were taken from 1-6-day old mice for primary culture. The morphology and the process of axon were observed by use of an inverted/phase contrast microscope. Immunocytochemical identification was performed using fluorescent microscopy. RESULTS: In the state of primary culture in vitro, the SGNs were found to have good cell morphology and axon regeneration power, the surviving cells of SGN(5-7 d) amounted to (78.10 +/- 4.33)/cm2 and the survival period was 7-14 d. CONCLUSION: We successfully established the methods of mechanical dissociation and culture of SGN cytological model; the cell amount and survival period of SGNs were able to meet the requirement of cytological experiments.