帕金森病脑内移植基因工程细胞系的构建  

The conformation of cell line by genetic engineering of intracerebral transplantation in PD

作　　者：杨扬[1] 王晓民[2] 吴承远[1] 卢峻[2] 韩济生[2] 

机构地区：[1]山东大学齐鲁医院神经外科,济南250012 [2]北京大学神经科学研究所

出　　处：《中华神经外科杂志》2002年第1期34-36,共3页Chinese Journal of Neurosurgery

摘　　要：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 human tyrosine hydroxylase (hTH)gene expressing cell-line for the experiment of gene therapy for Parkinson's disease(PD). Methods Recombinant plasmid pcDNA3/hTH was subcloned, purified and transfected into human neuroblastoma cell-line (hybridoma)-SH-SY5Y cells with liposome method (lipofectin). Under the pressure of G418(800μg/μl),drug-resistant clones were screened, separately cultured and assayed immunocytochemically to establish hTH expressing clones.Results SH-SY5Y cells transfected with pcDNA3/hTH were obviously stained immunocytochemically in contrast to the control SH-SY5Y cells which were merely nonspecifically stained.Conclusions SH-SY5Y cells transfected with pcDNA3/hTH can efficiently express hTH, which is potentially applicable to non-human primate experiments for the treatment of PD.

关 键 词：帕金森病 酪氨酸羟化酶 基因治疗 

分 类 号：R742.5[医药卫生—神经病学与精神病学]