A novel approach for gene transduction with adenovirus vector and the fibrin glue system.
BACKGROUND
The fibrin glue system (FGS) consists of liquid forms of fibrinogen and thrombin and is used widely in surgery for hemostasis. In this study, as a novel and unique approach, the possibility and efficacy of locoregional gene transfer using the FGS containing an adenoviral vector was determined.


METHODS
The optimum concentration of the adenoviral vector mixed with the FGS (AxCALacZ/FGS) for gene transduction was evaluated in vitro by X-gal (beta-galactosidase) staining and NIH (National Institute of Health) imaging in RCN-9, a rat colon carcinoma cell line. To determine the survival period of the adenoviral vector in the fibrin glue, RCN-9 cells were exposed to AxCALacZ/FGS after it had been incubated for various periods and the transduction efficiencies were evaluated by beta-galactosidase (beta-gal) assay. AxCALacZ/FGS was also applied in vivo to the resected site of rat liver. AxCALacZ diluted in PBS (AxCALacZ/PBS) was used as the control. The transduction efficiencies in the liver were compared by X-gal staining and beta-gal assay.


RESULTS
Almost 100% transgene expression was demonstrated by the X-gal staining and NIH imaging at a concentration level greater than 1 multiplicity of infection. LacZ expression (as beta-galactosidase) revealed gene-transduced RCN-9 cells when the AxCALacZ/FGS was held for a period of less than 96 hours. The treatment with the AxCALacZ/FGS in vivo resulted in greater transgene expression than the treatment with AxCALacZ/PBS.


CONCLUSIONS
The adenoviral vector survives and remains stable in the FGS for sufficient time for transduction to occur and AxCALacZ/FGS can efficiently transduce the target gene both in vitro and vivo.
Sakurai
Minori
M
Department of Surgery, the Jikei University School of Medicine, Tokyo, Japan.
Misawa
Takeyuki
T
Shiba
Hiroaki
H
Iida
Tomonori
T
Ohashi
Toya
T
Yanaga
Katsuhiko
K
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D015718:Fibrin Tissue Adhesive; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D014161:Transduction, Genetic / Q000379:methods*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics
D015718:Fibrin Tissue Adhesive; D001616:beta-Galactosidase