Gene transferred rat aorta grafting

AIM: To establish a rat sFGFR1 gene transferred aorta transplantation model for the study of accelerated graft arteriosclerosis (AGA) caused by chronic rejection. METHODS: ① sFGFR1 gene was cloned and expressed in RTS system. Adv sFGFR1 and Adv LacZ gene transfer vectors were constructed. ② Left renal arteries of donor rats were isolated and cannulated. The aortas below left renal artery and above iliac bifurcation were isolated and tied off. 100-200 μL (2×10 9-5×10 9 Pfu) Adv sFGFR1 or Adv LacZ was infused into aortas through cannula and the pressure was kept for 20 min. The aortas were then removed and orthrotopiclly transplanted into syngeneic recipient abdomen. The graft was removed 1 d, 3 d, 7 d, 14 d and 28 d post transplantation. Morphological changes, LacZ and sFGFR1 gene expression were analyzed by histological, β gal, histochemical and immunohistochemical staining and RT PCR examination. RESULTS: Target genes (sFGFR1 and LacZ ) were efficiently transfected into aorta endothelial cells and media smooth muscle cells by adenovirus vector mediated intra aorta infusion with isolated stable pressure. The expression of target gene was achieved up to 28 d post isografting. The level of target gene expression declined with the length of time. Morphological study revealed that sFGFR1 transfected aortas were normal in structure and no inflammation reaction was observed. CONCLUSION: Target gene (sFGFR1 and LacZ ) transfer can be safely and efficiently achieved by adenovirus vector mediated intra aorta infusion with isolated stable pressure. Gene transfected aortas can be grafted and maintain the target gene expression.