Recombinant Adeno-associated Virus Type 9 Transfection Of Mouse Bone Marrow Mesenchymal Stem Cells

Objective:To investigate the transfection efficiency of rAAV9-enhanced green fluorescence protein (eGFP) in transfection of mouse bone marrow mesenchymal stem cells.Methods:rAAV9-eGFP was transfected into mouse bone marrow mesenchymal stem cells which were subcultured to the fourth generation at different multiplicities of infection (MOI=1×105, 1×106, 1×107). To find the best condition of transfection, we observed the eGFP expression under the inverted fluorescence microscope at 1-7 days after transfection. Then the transfection efficiency of rAAV9-eGFP under the optimal MOI was determined by flow cytometry.Results:The cells with rAAV9-eGFP transfection at MOl of 1×107 began to exhibit eGFP expression at the 1st day after transfection, and the cells transfected at MOI of 1×105 and 1×106 began to exhibit eGFP expression from the 2nd day after transfection. The eGFP expression increased with the MOI used for transfection, and the intensity of eGFP expression increased with the time going on. At the 5th day, the eGFP expression reached the peak value, and meanwhile, the transfection efficiency at MOI=1×107was about 8%. Conclusion:rAAV9-eGFP cannot transfect mouse bone marrow mesenchymal stem cells effectively.