In vivo tracking of adipose-derived stem cells labeled with green fluorescent protein and luciferase dual reporter gene

Abstract

Objective T o explore the feasibility of in vivo tracking of adipose-derived stem cells( ADSC s) by using lentiviral vectors containing green fluorescent protein( GFP) and luciferase( Luc) dual reporter gene transfection and bioluminescent imaging techniques. Methods T he ADSC s were isolated from human adipose tissue samples of cosmetic subdermal liposuction. T he ADSC s were transfected by lentiviral vectors containing GFP and Luc. T he labeling efficiency was evaluated by fluorescence microscope and flow cytometry in vitro. T he transfected ADSC s were subcutaneously injected into BALB /C mice, and then a non-invasive bioluminescence imaging procedure was performed 1 h, 1, 2, 4, 6 and 8 weeks after transplantation. T he DAPI staining of skin tissues was performed to identify the survival and the location of transplanted ADSC s. Results High GFP expression was observed in transfected ADSC s by lentiviral vector under fluorescence microscope. T he transfection efficiency of lentiviral vectors containing GFP and Luc was 87. 3%. T ransfected ADSC s can express GFP and Luc after 25 successive generations, and the transduction of ADSC s with lentiviral bioluminescence reporter vectors had no effect on cell proliferation. T he bioluminescence of ADSC s in vivo can be detectable as long as 8 weeks after transplantion, and the ADSC s mainly survive in the inoculating site of the thigh.Histological analysis of sections showed that the labeled ADSC s located in the subcutaneous layer of the implantation site. Conclusion U sing lentiviral vectors containing GFP and Luc dual reporter gene transfection and bioluminescent imaging techniques can monitor ADSC s in vivo.