摘要
目的 通过神经营养素 3(NT3)基因转染的方法 ,了解NT3对耳蜗螺旋神经节细胞(SGC)生长的影响。方法 建立体外培养小鼠耳蜗螺旋神经节细胞的方法 ,并进行神经丝蛋白 (NF)免疫组化染色鉴定及受体酪氨酸激酶C(TrKC)免疫组化染色。利用阳离子脂质体作为载体 ,将重组质粒 pIRES2 EGFP(增强型绿色荧光蛋白 ) NT 3瞬时转染耳蜗螺旋神经节细胞 ,观察转染后耳蜗螺旋神经节细胞的形态、数量等改变。结果 成功培养了小鼠耳蜗螺旋神经节细胞。NT3基因转染 4 8h后 ,荧光显微镜下见聚合分布的、胞体及轴突发出绿色荧光的螺旋神经节细胞 ,与未转染细胞相比 ,螺旋神经节细胞形态未见明显变化。继续培养 2周后 ,各实验组细胞的数量均减少 ,但NT3转染组细胞减少的数量低于空载体组及空白对照组。
Objective To observe the influence of neurotrophin-3 (NT3) transfection in vitro on the growth of mice cochlear spiral ganglion cells(SGCs).Methods The SGCs were cultured and purged in vitro. SGCs were identified by neurofilament (NF) and receptor tyrosine kinase C (TrKC) immunocytochemical staining. The pIRES2-EGFP(enhanced green fluorescent protein)-NT3 was transfected into SGCs by using lipofectamine. The SGCs with green fluorescence were observed by fluorescence microscope. The number of SGCs from different groups was measured after 2 weeks.Results The SGCs with green fluorescence were successfully observed under fluorescence microscope. The decreasing number of SGCs in the group transfected with NT-3 was less than that in the control groups.Conclusion The degradation of the cochlear spiral ganglion cells can be alleviated by NT3 gene transfection in vitro.
出处
《中华神经外科疾病研究杂志》
CAS
2004年第3期249-251,共3页
Chinese Journal of Neurosurgical Disease Research
