INFECTION OF NEURAL STEM CELLS WITH RECOMBINANT RETROVIRUSES CONTAINING NGF OR GDNF cDNA AND THEIR IDENTIFICATION / 解剖学报

ABSTRACT

Objective To explore whether the neural stem cells(NSCs) can act directly as a gene target cell which can be infected by the recombinant retrovirus and express the products of exogenous genes after infection. Methods The NSCs were cultured with supernatant containing the recombinant retroviruses with the genes of NGF or GDNF for two days.After screened with G418,the infected NSC were expanded at the present of bFGF in culture.The PC12 cells and the neurons of ventral midbrain of rat were cultured by the medium from the infected NSC,which were called as GDNF\|containing conditioned medium NGF or GDNF\|containing conditioned medium the morphological changes of the dopamine neurons of the ventral midbrain and expression of exogenous genes of the infected NSCs were detected by immunohistochemistry staining. Results It was estimated that about fifty percent of NSCs via retrovirus\|mediated NGF or GDNF gene transduction were G418\|resistant.These infected NSCs began to differentiate.Long and radical processes reached out from the sphere of proliferation and the cells migrated towards outside along the processes.The NSC infected with gene of NGF showed an astroid\|shape with larger body and processes.The NSC infected with gene of GDNF showed a shuttle\|shape with a smaller body and long processes.The PC12 cells increased in the NGF\|containing conditioned medium and stretched out long neurites.The dopamine neuron of the ventral midbrain which were immunoreactive for TH also showed a larger body and longer processes in the GDNF\|containing conditioned medium.Most of G418\|resistant NSCs were immunoreactive for NGF or GDNF. Conclusion NSC can act directly as a gene target cell which not only be infected by the recombinant retrovirus,but also express and secrete the products of exogenous genes.