RRC ID 43541
Author Sakurai M, Misawa T, Shiba H, Iida T, Ohashi T, Yanaga K.
Title A novel approach for gene transduction with adenovirus vector and the fibrin glue system.
Journal Anticancer Res
Abstract 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.
MATERIALS AND 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.
CONCLUSION: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.
Volume 28(6A)
Pages 3809-13
Published 2008-1-1
PMID 19189668
MeSH Adenoviridae / genetics* Animals Cell Line, Tumor Fibrin Tissue Adhesive* Gene Transfer Techniques* Genetic Vectors / genetics* Male Rats Rats, Inbred F344 Transduction, Genetic / methods* beta-Galactosidase / analysis beta-Galactosidase / biosynthesis beta-Galactosidase / genetics
IF 1.994
Times Cited 0
Human and Animal Cells RCN-9(RCB0511)