Comparison Of Lentiviral Vector And Adenoviral Vector Mediated EGFP/BMP-2 Transfer Into Rabbit Bone Mesenchymal Stem Cells

Objective To compare the duration,efficiency and the deviation of exogenous gene expression of transfection of rabbit bone mesenchymal stem cells(BMSCs)using lentiviral vector and adenoviral vector which mediated enhanced green fluorescent proteins(EGFP)and bone morphogenetic protein2(BMP-2).Method BMSCs were isolated with whole bone marrow culture method and purified using attachment method.Surface marks were detected by flow cytometry.The 5th generation of rabbit BMSCs were exposed to Ad-EGFP-BMP-2(group A)and Lenti-EGFP-BMP-2(group B)with multiplicity of infection(MOI)of 100,as transfection groups.And in contrast group(group C),the same quality of culture medium was request equivalent to the group A and B.The expression of EGFP was observed by inverted fluorescence microscope at various time intervals.And the expression of exogenous gene BMP 2 in cell was detected and analyzed by immunohistochemical staining and western blot after transfection.Result The 5th generation of rabbit BMSCs with high purity were successfully obtained.Flow cytometry analysis showed that CD44,CD29 were positive while CD45 was negative.The intense green fluorescence emerged under the microscope in 24~48h after transfected in group A,which is earlier than group B.High expression of BMP-2 was found by western blot also by immunohistochemical staining and was downregulated after 1 week in group A.Group B showed the different,high expression of EGFP/BMP-2 persisted for a longer period after transfected even lasted for 3 weeks.Conclusion The lentiviral vector and adenoviral vector can efficiently transfected rabbit BMSCs and stably express the target gene of EGFP/BMP-2.Under the same MOI,compared to adenoviral vector,transfection of lentiviral vector to rabbit BMSCs is more effectively and expression of EGFP/BMP-2 can be persistent in a longer term.