Compare The Three Methods With The Carried IDO Adenoviral Transfected Liver Cells

Objective: To study three kinds of ways of carrying transfected IDO (indoleaminc2,3-dioxygcnasc,IDO) adenovirus transfection efficiency of mouse liver cells, liver cells, IDO activity and geneexpression.METHODS: The recombinant adenovirus which carrying the same dose IDO transfect the BABL/cmouse liver in three ways.using collagenase perfusion in vivo and in vitro digestion method toisolated mouse liver cells, trypan blue dye assess liver cell activity, and using RT-PCR and WesternBlot were detected after transfection of liver cells in IDO gene and protein expression.Results: The portal vein infusion centrifugation can successfully separated and purified liver cells,liver cells1.62×10~8/only. Adenovirus can successfully transfected liver cells, the averagetransfection rate of the three groups were: in vitro group27.298%,79.076%activity of liver cells.Portal group24.676%,94.768%activity of liver cells.21.01%of peripheral intravenous group, livercell activity is90.812%. RT-PCR, WestBlot can detect that IDO gene can successful express in livercells.Conclusion: The portal vein injection group can get high transfection efficiency, high activity of livercells.