|Budget Amount *help
¥12,400,000 (Direct Cost : ¥12,400,000)
Fiscal Year 1999 : ¥3,500,000 (Direct Cost : ¥3,500,000)
Fiscal Year 1998 : ¥8,900,000 (Direct Cost : ¥8,900,000)
(1) Ex vivo gene transfer with gene-gun
We evaluated gene-gun-mediated ex vivo gene transfer using subcuateous tumor-bearing rat model. First we transplanted 9L glioma cells into F344 syngeneic rats subcutaneously and surgically removed formed tumors 20 days after transplantation. Lac Z expression vector was transduced by gene-guun at shooting pressure (psi) 100, 200, 400, and 600 (max) or Lca Z expressing adenovirus was incubated with the tumor at MOI 20, 40, and 100 for 24 hours. The tumors were then re-transplanted subcutaneously and gene expression was evaluated 7 days after the re-transplantation. The highest gene-gun-mediated gene transfer rate ranged from 3 to 9 % along with shooting pressure whereas that by adenovirus ranged from 5.5 to 15.9% along with MOI which are statistically different.
(2) In vivo gene transfer with gene-gun
Next, we estimated gene-gun mediated in vivo gene transfer using the same animal model as (1). We surgically opened the skin over the subcutaneously formed tumors and shot Lac Z expression vector at the same psi as (1) or injected Lac Z expression adenovirus at the same MOIs as (1). After the treatments, the wounds were closed and gene expression was examined 7 days after the treatments. The highest gene-gun-mediated gene transfer rate ranged from 3.6 to 9.5 % along with shooting pressure whereas that by adenovirus ranged from 5.8 to 13.2 % along with MOI which are statistically different.
In conclusion, although gene transfer with gene-gun is less effective than adenovirus-mediated gene transfer against gliomas, gene-gun-mediated gene transfer is still a good tool to transduce genes in vivo or ex vivo.