TH基因修饰的中脑星形胶质细胞的建立

目的 体外构建酪氨酸羟化酶(TH)基因修饰的中脑星形胶质细胞，为进行帕金森病(PD)的转基因细胞治疗奠定基础。方法 应用逆转录病毒表达载体将鼠TH基因转入逆转录病毒包装细胞PA317，通过G418抗性筛选，获取高滴度产毒PA317／pN2ATH细胞株，以其产生的病毒上清感染鼠中脑星形胶质细胞，继续培养7～9d后，免疫组化检测细胞TH表达。结果获得了高滴度产毒PA317／pN2ATH细胞，并且转染pN2ATH的中脑星形胶质细胞成功表达TH。结论转染pN2ATH的中脑星形胶质细胞可以有效表达TH，为将其应用于PD基因治疗实验研究提供了基础。

Construction of Tyrosine Hydroxylase Gene Modified Mesencephalic Astrocytes

Objective To construct tyrosine hydroxylase(TH) gene expressing mesencephalic astrocytes for the experiment of gene therapy for Parkinson's disease(PD). Methods The recombinant retroviral vector pN 2ATH was constructed and transferred into PA317 pakaging cells by electroporation method.Resistant clones were obtained with G418 selective culture, and the virus supernatants were harvested. The mesencephalic astrocytes were cultured with the pN 2ATH viral supernants. After 7-9 days culture,TH expression in the infected astrocytes were detected by immunohistochemistry. Results After G418 selection, high titer PA317/pN 2ATH cells were obtained. The mesencephalic astrocytes transfected with pN 2ATH succesfully expressed TH. Conclusion Mesencephalic astrocytes transfected with pN 2ATH can efficently express TH,which may provide a basis for further studies on gene therapy of PD in animals.