Table 1.
Tumor take rates in different models.
Fig 1.
Systemic injection of Hepa1-6 cells in syngeneic C57BL6 mice.
Hepa1-6 cells (2x106) suspended in PBS were injected into C57BL6 mice intravenously. (a) Morphological examination of tumor nodules in different tissues. The results showed that tumor formation in lung, heart and ribs. (b) Histological assessment of liver tumor nodules in lung, liver and heart (scale bar = 50 μm). A’, B’ or C’ represents the corresponding magnified boxed area from A, B or C.
Fig 2.
Intrasplenic and intrahepatic inoculation of Hepa1-6 cells in C57BL6 mice.
Hepa1-6 cells (2x106) suspended in PBS were injected into C57BL6 mice intrasplenicly or intrahepatically as described in Materials and Methods. (a) Morphological examination of tumor nodules in different tissues from orthotopic HCC mice generated by intrasplenic inoculation of Hepa1-6 cells. The results showed the tumor formation in liver and spleen. (b) Histological assessment of liver tumor nodules in spleen, liver and lung (scale bar = 100 μm). A’, B’ or C’ represents the corresponding magnified boxed area from A, B or C. (c) MRI analysis of the progression of liver tumors after intrahepatic inoculation of Hepa1-6 cells at different time-points. Arrows point to the tumor nodules. (d) Morphological examination of tumor nodules in liver from orthotopic HCC mice via intrahepatic injection of Hepa1-6 cells. The results showed both solitary and multinodular tumors formed in liver. (e) Histological assessment of liver tumor nodules in liver and lung (scale bar = 100 μm). A’, B’ or C’ represents the corresponding magnified boxed area from A, B or C.
Fig 3.
Intrahepatic tissue implantation in C57BL6 mice.
Hepa1-6 cells (3x106) were suspended in 50μl PBS and injected into the left axilla of C57BL6 mice subcutaneously with 1 ml syringe. Subsequently, tumor tissues were cut into about 1mm3 pieces. Tumor pieces with a size about 1mm3 were implanted in the left liver lobe of C57BL6 mice. (a) MRI analysis of the progression of liver tumors after tissue implantation at different time-points. Arrows point to the tumor nodules. (b) Measurement of liver tumor sizes at different time-points after implantation. The data represents as mean±sem and significant difference was detected between different time-points (*p<0.05; **p<0.01, n = 10 for 2 week time points; n = 20 for 3 week time point; n = 19 for 7 week time point after implantation). (c) Assessment of survival rates of orthotopic HCC mice generated by intrahepatic tissue implantation (n = 19). (d) Morphological examination of tumor nodules in different tissues. The results showed the tumor formation in liver, peritoneum, mesenterium and diaphragm. (e) Histological assessment of liver tumor nodules in lung and liver (scale bar = 100 μm). A’, B’ or C’ represents the corresponding magnified boxed area from A, B or C. (f) Immunohistochemistry of CD3+ and Foxp3+ regulatory T cells in liver sections from orthotopic HCC mice and HCC patients to determine the extent of T cell infiltration (scale bar = 100 μm). Arrowhead points to CD3+ or Foxp3+ T cells. TI represents Tissue Implantation; CI denotes as Cell Inoculation. (g) Measurement of immune cytokines including IFN-γ and IL-10 in serum from orthotopic HCC mice (n = 5, *P<0.05; **P<0.01). The comparison was conducted between CI, TI and normal controls. TI represents Tissue Implantation; CI means Cell Inoculation.