[酪氨酸羟化酶基因修饰中脑星形胶质细胞的构建]
[Construction of tyrosine hydroxylase gene modified mesencephalic astrocytes].
作者信息
Xie Xuan-gui, Liu Jun, You Chao, Yang Hui, Zhang Ke-cheng
机构信息
Department of Neurosurgery, West China Hosiptal, Sichuan University, Chengdu 610041, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2004 Sep;35(5):603-6.
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 pN2ATH 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 pN2ATH viral supernants. After 7-9 days culture, TH expression in the infected astrocytes were detected by immunohistochemistry.
RESULTS
After G418 selection, high titer PA317/pN2ATH cells were obtained. The mesencephalic astrocytes transfected with pN2ATH succesfully expressed TH.
CONCLUSION
Mesencephalic astrocytes transfected with pN2ATH can efficently express TH,which may provide a basis for further studies on gene therapy of PD in animals.
目的
构建表达酪氨酸羟化酶（TH）基因的中脑星形胶质细胞，用于帕金森病（PD）的基因治疗实验。
方法
构建重组逆转录病毒载体pN2ATH，采用电穿孔法将其转入PA317包装细胞。通过G418选择性培养获得抗性克隆，收集病毒上清液。用pN2ATH病毒上清液培养中脑星形胶质细胞。培养7 - 9天后，通过免疫组织化学检测感染星形胶质细胞中TH的表达。
结果
经G418筛选后，获得了高滴度的PA317/pN2ATH细胞。用pN2ATH转染的中脑星形胶质细胞成功表达了TH。
结论
用pN2ATH转染的中脑星形胶质细胞能有效表达TH，这可能为进一步开展PD动物基因治疗研究提供基础。
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