Transfection of rat bone marrow mesenchymal stem cells with adenovirus vector carrying hBcl-2 and hVEGF_(165) genes.

Objective Bone marrow mesenchymal stem cells(BMSCs) were transfected using recombinant adenovirus vector carring hBcl-2/hVEGF165 co-expression genes for assessment of expression and biological effects of these.Methods Rat BMSCs were isolated and purified by attachment culture methed,subsequently transfected with recombinant adenovirus vector.Targeted mRNA expression was quantified by PCR.Western Blot and Elisa were applied to assess protein expression of Bcl-2 and hVEGF165 in transfected cells.Finally,the biological effects of hVEGF165 and Bcl-2 in were observed via MTT and apoptosis detection.Results Bone marrow mesenchymal stem cells were separated and purified and transfected by recombinant adenovirus successfully.The mRNA of hBcl-2 and hVEGF165 were detected after transfection.hVEGF165 in the supernatant of transfected cells was detected,with peak concentration of(924.3±56.5)pg/mL,by Elisa Kit,with significant differences comparing to those in control group in at different time spots(P0.05).Meanwhile,significant increase of Bcl-2 protein was detected in transfected group by Western(P0.05).Proliferation of BMSCs was significantly promoted with hVEGF165-containing supernatant using MTT assay(P0.05).Significant reduction of apoptosis was also observed in transfected BMSCs using annexin V-PE/7-AAD detection(P0.05).Conclusion Transfected BMSCs could express functional target genes,hBcl-2 and hVEGF165.