Construction of Retroviral Vector Encoding Human Insulin-like Growth Factor-1 and Its Expression in Bone Marrow Stromal Cells

Objective To construct a retroviral mediated expression system of human insulin-like growth factor-1(IGF-1),and determine whether bone marrow stromal cells(BMSCs) can be infected by retrovirus.Methods hIGF-1cDNA was amplified in vitro from normal human liver cells by using RT-PCR and pLXSN was cloned into plasmid vector.The recombinant vector pLXSN/IGF and control vector pLXSN were transfected into packaging cell PA317 and G418 was used to select positive colony.Hu-man BMSCs cells were infected with a high titre viral supernatant.The expression of hIGF-1 in target cells were analyzed with RT-PCR assay and Western blot.Results The retroviral vector containing IGF-1 gene was constructed successfully,amphotropic PA317 retroviral producer cells were obtained by liposome transfection and G418 selelction.The BMSCs cells were infected by medium containing IGF-1 virus.The expression of IGF-1 in transfected BMSCs was dectected by RT-PCR and Western blot analysis.Conclusion The constructed retrovirus vectors can introduce IGF-1 into BMSC cells efficiently,which may have potential effect for gene therapy of bone repair.