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目的 为进行帕金森病 (PD)的转基因细胞治疗 ,在实验室构建人类酪氨酸羟化酶(hTH)基因表达工程细胞系。方法 重组质粒pcDNA3/hTH经亚克隆、纯化后 ,用脂质体法转染到人类神经母细胞瘤细胞系SH SY5Y细胞内。经G418筛选、克隆 ,分离培养经免疫细胞化学分析。结果 转染pcDNA3/hTH的SH SY5Y细胞免疫细胞化学染色明显 ,与仅呈非特异性染色的对照组SH SY5Y细胞形成鲜明的对比。结论 转染pcDNA3/hTH的SH SY5Y细胞能够有效地表达hTH ,在PD治疗的非人类灵长目实验中具有潜在的应用价值。
Objective To construct a human tyrosine hydroxylase (hTH) gene expression engineering cell line in the laboratory for the treatment of transgenic cells with Parkinson’s disease (PD). Methods The recombinant plasmid pcDNA3 / hTH was subcloned and purified. The recombinant plasmid pcDNA3 / hTH was transfected into human neuroblastoma cell line SH SY5Y by liposome. After G418 selection, cloning, isolation and culture by immunocytochemistry. Results The SH SY5Y cells transfected with pcDNA3 / hTH showed obvious immunocytochemical staining, which was in striking contrast with the SH SY5Y cells in the non-specific staining. Conclusions SH SY5Y cells transfected with pcDNA3 / hTH can effectively express hTH, which has potential value in the non - human primate test of PD.