The experimental study of genetic engineering human neural stem cells mediated by lentivirus to express multigene
Pei-qiang Cai1, Xun Tang, Yue-qiu Lin
1Department of Orthopaedics, First People's Hospital of Yibin, Yibin 644000, China. cpq20032002@yahoo.com.cn
Objective:
To explore the feasibility to construct genetic engineering human neural stem cells (hNSCs) mediated by lentivirus to express multigene in order to provide a graft source for further studies of spinal cord injury (SCI).
Methods:
Human neural stem cells from the brain cortex of human abortus were isolated and cultured, then gene was modified by lentivirus to express both green fluorescence protein (GFP) and rat neurotrophin-3 (NT-3); the transgenic expression was detected by the methods of fluorescence microscope, dorsal root ganglion of fetal rats and slot blot.
Results:
Genetic engineering hNSCs were successfully constructed. All of the genetic engineering hNSCs which expressed bright green fluorescence were observed under the fluorescence microscope. The conditioned medium of transgenic hNSCs could induce neurite flourishing outgrowth from dorsal root ganglion (DRG). The genetic engineering hNSCs expressed high level NT-3 which could be detected by using slot blot.
Conclusions:
Genetic engineering hNSCs mediated by lentivirus can be constructed to express multigene successfully.
