Construction of EGFP-labled recombinant plasmid of VEGF165 gene and its expression in bone marrow mesenchymal stem cells / 解放军医学杂志

ABSTRACT

Objective To construct a EGFP-labled recombinant plasmid of VEGF165 (vascular endothelial growth factor) gene, and to study the transfection and expression of VEGF165 eukaryotic expression plasmid in mesenchymal stem cells (MSCs). Methods pIRES2-EGFP-VEGF165 recombinant plasmid was constructed, which was then transfected into rat MSCs. ELISA and MTT were used to detect the expression level and biological activity of VEGF in the conditioned medium after transfection. Results There was a significant increase in VEGF protein in the MSCs after being transfected with pIRES2-EGFP-VEGF165. The conditioned medium after transfection showed the biological activity of stimulating the proliferation of endothelial cells. Conclusions The pIRES2-EGFP-VEGF165, a eukaryotic expression plasmid for VEGF165 gene, is constructed. High levels of VEGF protein expression can be obtained in the MSCs transfected with pIRES2-EGFP-VEGF165. The expressed protein has the biological activity of VEGF.