Nerve growth factor (NGF) belongs to a biological macromolecule that is difficult to pass through the blood-brain barrier. However, a retroviral vector carrying exogenous gene can be stably inserted and integrated into the host cell genome, which is suitable for gene therapy.OBJECTIVE:
To study the gene expression of recombinant retroviral vector carrying rat NGF gene in neural stem cells.METHODS:
The rat NGF gene was inserted into a retroviral vector pLEGFP-N1, which was transferred into packaging cells PT67 by Lipofectamine 2000. The positive clones in virus supernatant were collected by G418 selection and used to infect neural stem cells. After that, the NGF expression was tested by enzyme linked immunosorbent assay and the biological competence by PC12 cells, and then morphological change of neural stem cells and cell typing were examined by fluorescent microscope.RESULTS AND CONCLUSION:
The neural stem cells could express extrinsic source NGF protein after the recombinant plasmid was infected into neural stem cells. The PC12 cells increased in the experimental group and stretched out long neurites. And the NGF protein could maintain the neural stem cell survival and stimulate their differentiation. These findings suggest that the neural stem cells carrying extrinsic source NGF gene could express NGF successfully, and the NGF protein induced the survival and differentiation of neural stem cells.RESULTS AND CONCLUSION:
Funding: the Natural Science Foundation of Guangdong Province, No. 04009566; the Medical Research Fund of Guangdong Province, No. A2003276; the Science and Technology Plan of Guangzhou City, No. 041011RESULTS AND CONCLUSION:
He JF, Gu GM, Long DH. Expression of recombinant retroviral vector carrying nerve growth factor gene in neural stem cells. Zhongguo Zuzhi Gongcheng Yanjiu. 2014;18(19):3100-3104.