Research On Osteogenic Characteristics Of Adipose-derived Mesenchymal Stem Cells Modified By Human VEGF Gene

Objective: To study the role of human VEGF gene in inducing osteogenic differentiation of adipose-derived mesenchymal stem cells after transfection with lentivirus.Methods: Adipose-derived mesenchymal stem cells were subcultured in vitro and stored in a cell incubator.The VEGF gene sequence was searched in Gene Bank,and the target fragment of interest gene was amplified by PCR.A lentiviral vector carrying human VEGF gene was proliferated in 239 T cells and purified afterwards,and the virus titer were detected.The subcultured adipose-derived mesenchymal stem cells were evenly divided into three groups:VEGF-CD513B-1,CD513B-1,and normal cells,and the recombinant lentiviruses,unrecombined lentiviruses,and normal saline were added and co-cultured with cells in the three groups,respectively.The activity of adipose-derived mesenchymal stem cells and the VEGF gene expression were detected after culture.Then osteogenic induction was performed on adipose-derived mesenchymal stem cells,and alizarin red staining was used to examine the formation of bone nodules to evaluate the osteogenic induction of VEGF gene on adipose-derived mesenchymal stem cells.Results: After lentivirus transfection,the proliferation rate of adipose-derived mesenchymal stem cells slowed down and took a little longer to reach the plateau.The final cell number was comparable to the normal h AMSC.After h AMSC osteogenesis induction,the gene expression and protein levels of VEGF and BMP2 were increased in the experimental group,and the increase was not significant.After the osteogenesis induction,alizarin red staining showed that bone nodules were formed in each group,but the difference was obvious.Bone nodule formation was significantly higher in VEGF recombinant adenovirus transfected adipose-derived mesenchymal stem cell group than in the other two groups.Conclusion: VEGF has significant osteogenic induction effects on adipose-derived mesenchymal stem cells,and the osteogenic effect of adipose-derived mesenchymal stem cells can be significantly improved by lentiviral transgenic technology.