Construction of Recombinant Adenovirus Co-expressing BMP9,BMP7 and Effect on Osteogenesis of Mesenchymal Stem Cells C3H10

Objective To construct a recombinant adenovirus co-expressing BMP9 and BMP7,and identify its primary effect on osteogenesis.Methods The gene sequence of BMP9 and BMP7 were amplified from AdEasy vector by PCR and sub-cloned into pASG2 vector.The co-expression shuttle plasmid pASG2-BMP9,7 was confirmed by restriction endonuclease digestion,PCR and gene sequencing;then plasmid pAdeasy-1 was co-transfected to produce recombinant adenovirus plasmid by homologous recombination.In addition,the recombinant vector was transfected into HEK293 cells.High-titer recombinant adenovirus(Ad-BMP9,7) was collected after rounds of amplification.The capability of osteogenesis induced with Ad-BMP9,7 was evaluated in vitro in C3H10 cells by alkaline phosphatase staining and calcium-alizarin red staining.Results Ad-BMP9,7 was constructed successfully.Testing with C3H10 cells,alkaline phosphatase staining and calcium-alizarin red staining were stronger than single transfected BMP7 or BMP9 cells.Conclusion The recombinant adenovirus co-expressing BMP9 and BMP7 was constructed successfully,which has a capability of inducing osteogenic differentiation of C3H10 cells.