Controllable HGF Expression Of BM-MSC Through Baculovirus System

Objective:To construct an efficient and regulable recombinant baoulovirus vAcrtTA2s-Ptight-HGF and investigate the optimum dose of deoxycycline(DOX) for the induction of hepatocyte growth factor(HGF) expression of bone marrow mesenchymal stem cells(BM-MSCs) infected with vAcrtTA2s-Ptight-HGF.and observe the changes of BM-MSCs morphology and proliferationrate after transfection.Methods:vAcrtTA2s-Ptight-HGF was built by gene recombination. BM-MSCs of New Zealand white rabbits were isolated and cultured by adhesion method and then were infected with vAcrtTA2s-Ptight-HGF The concentration of HGF secreted by infected rabbit RM-BM-MSCs was verified by ELISA and Western with differernt induction dose of DOX.Results:Rabbit BM-MSCs of the fifth generation was successfully cultured. An efficient and regulable recombinant baoulovirus vAcrtTA2s-Ptight-HGFwas successfully constructed by gene recombination and infect rabbit BM-MSCs. Different levels of HGF expression at different doses of deoxycycline were quantitatively verified by ElISA and Western blot and for7consecutive days. And the amount of HGF expression in the experimental group was significantly higher than the control group (P<0.001), transfected BM-MSCs were still scrollshaped, Serial concentrations of HGF promote BM-MSCs proliferation rate was significantly higher than that in control group (P<0.001).Conclusion:Recombinant baculovirus vAcrtTA2s-Ptight-HGF was successfully constructed, and realize the controllable expression of HGF, the optimum dose of DOX to induce HGF expression in BM-MSCs in vitro was1μg/ml. and confirmed that HGF can promote the proliferation of BM-MSCs,transfected BM-MSCs morphology did not change significantly.