Controllable HGF expression in bone marrow mesenchymal stem cells through baculovirus system

Abstract:

Objective：To construct an efficient and controllable recombinant baoulovirus vAcrtTA2s-Ptight-HGF and to investigate the optimum dose of deoxycycline（DOX） for the induction of hepatocyte growth factor（HGF） expression in bone marrow mesenchymal stem cells（BM-MSCs） infected with vAcrtTA2s-Ptight-HGF. Methods：vAcrtTA2s-Ptight-HGF was built by gene recombination. BM-MSCs of New Zealand white rabbits were isolated and cultured by adhe－sion method and then were infected with vAcrtTA2s-Ptight-HGF. Concentration of HGF secreted by infected rabbit BM-MSCs was verified by ELISA and Western blot with different induction doses of DOX. Results：An efficient and controllable recombinant baoulovirus vAcrtTA2s-Ptight-HGF was successfully constructed by gene recombination and rabbit BM-MSCs was successfully infected. Different levels of HGF expression under different doses of DOX were quantitatively verified by ELISA and Western blot. Expression of HGF was in dose-dependent relationship with DOX doses and was consistently lasted for 7 d. Expression of HGF was higher in experiment group than in control group（P<0.001）. Conclusions：BM-MSCs/vAcrtTA2s-Ptight-HGF demonstrates higher HGF expression and the optimum dose of DOX to induce HGF expression in infected rabbit BM-MSCs was 1 μg/ml.