In vitro study of transfection of mesenchymal stem cells with adenoviral vector overexpressing human uPA / 上海第二医科大学学报

ABSTRACT

Objective To identify the isolated rat bone marrow derived mesenchymal stem cells(MSCs),and evaluate the efficiency of adenoviral vector expressing human urokinase type plasminogen activator(uPA) in transfection of rat MSCs and its effect on proliferation of MSCs. Methods MSCs were isolated and purified by pasted wall purification,and were identified by immunicytochemistry.The transfection efficiency of uPA was detected by fluorescent microscopy,the expression of uPA in MSCs was detected by Western blotting,and the proliferation of MSCs was evaluated by MTT. Results The harvested MSCs exhibited the typical appearance of MSCs,and it was revealed by immunohistochemistry that the expression of MSCs markers CD29 and CD90 was positive,while that of CD34 and CD45 was negative.A tendency of increase in expression of green fluorescent protein(GFP) was observed with increase of multiplicity of infection(MOI).After transfection with AduPA for 72 h,the transfection efficiency reached(94.0?1.5)% at MOI of 80,and positive GFP cells could still be observed even after 7 d.The transfected uPA had no effect on the proliferation of MSCs. Conclusion MSCs are favourable genetic vectors to express uPA,and can be used for treatment of liver fibrosis.