Experimental Study On BFGF Gene Lentiviral Vector Transfecting BMSCs

Objective To enhance the expression of basic fibroblast growth factor (bFGF)in bone mesenchymal stem cells(BMSCs),explore the ways of bFGF and green fluorescent protein(GFP)gene highly transfected to BMSCs by lenti-bFGF-GFP and the efficiency of different MOI(multiplicity of infection)transfected to BMSCs.Methods BMSCs were separated and purified in vitro from whole bone marrow.BMSCs was subject to different treatments with three groups,that was bFGF-GFP-BMSCs group, GFP-BMSCs group and BMSCs group.72 hours after culture of BMSCs,ELISA and RT-PCR was adopted to quantify the amount of protein and mRNA of bFGF in the different groups.lentivirus with different MOI(10,30,50)was transfected into BMSC and GFP expression was examined with fluorescence microscope.Results The results of ELISA experiment discovered this:72 hours after transfection,the amount of bFGF protein of bFGF-GFP-BMSCs group was much higher than these of the GFP-BMSCs group and BMSCs group(P<0.01).RT-PCR showed that: compared to the GFP-BMSCs group and BMSCs group,the RNA content of bFGF in bFGF -GFP-BMSCs group was significantly increased after incubated 72 hours.When the MOI equal to 10,30,50,the efficiency of lentivirus transfected into BMSCs was 36.6%,84.2%,98.5%.conclusion Lentivirus mediated by bFGF and GFP gene can infect BMSCs successfully and which can express and secret bFGF protein after transfection.BMSCs infected by lentivirus which was mediated by bFGF and GFP gene is an efficient way for gene therapy of spinal cord injury (SCI).When MOI equal to 50,the efficiency exceeded 85%.