Expression of Vascular Endothelial Growth Factor after Transfection of Human Neural Stem Cells with the Lentiviral Vector Encoding the VEGF165 Gene

We examined the expression of vascular endothelial growth factor (VEGF) and characteristics of human neural stem cells after transfection with the lentiviral vector encoding the VEGF 165 gene. The latter gene was amplified from the human breast cancer cell line MCF-7 using RT-PCR; then the target gene was cloned into the pCDH–CMV–MCS-EF1-copGFP, a lentiviral expressing plasmid. After transformation, enzyme digestion led to a correct length of the VEGF165 gene, and DNA sequencing analysis confirmed that the VEGF165 gene sequence was exactly the same as that reported by the GeneBank. Then, the recombinant lentivirus produced by 293T cells and packaging plasmids were transfected into fourthpassage human neural stem cells (NSCs). One week after transfection with pCDH-VEGF165, NSCs expressed VEGF stably, and their proliferation ability significantly increased. Furthermore, human NSCs kept their characteristics and multiple differentiation activity after transfection. Our results indicate that human NSCs can express vascular endothelial growth factor highly and stably via transfection with the lentiviral vector encoding the VEGF165 gene, which may be useful for future research on function recovery after stroke.

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