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.