Figure 3.

Influence of the interaction between tumor-associated neutrophils (TANs) and tumor-associated macrophages (TAMs) on intrahepatic cholangiocarcinoma (ICC) growth and metastasis. (A) Schematic of the mouse xenograft experiments. Control group: 1×10⁷ ICC cells were injected into the subcutaneous space of the upper left flank region of NOD-Prkdc^scid IL2rg^tm1/Bcgen mice at day 0. TANs group: 1×10⁷ ICC cells were co-injected with 1×10⁶ TANs into the subcutaneous space of the upper left flank region of mice at day 0, and TANs were injected into the tumor at the indicated time (red arrows). TAMs group: 1×10⁷ ICC cells were co-injected with 1×10⁶ TAMs into the subcutaneous space of the upper left flank region of mice at day 0, and TAMs were injected into the tumor at the indicated time (yellow arrows). TANs+TAMs group: 1×10⁷ ICC cells were co-injected with 1×10⁶ TANs and TAMs mixture (TANs:TAMs=1:1) into the subcutaneous space of the upper left flank region of mice at day 0, and TANs and TAMs mixture were injected into the tumor at the indicated time (blue arrows). All mice were monitored once every 5 days and killed 5 weeks later. (B) Xenografts containing TANs and TAMs produced greater tumor volume and more pulmonary metastasis (C) than xenografts composed of ICC cells alone, *p<0.05, **p<0.01, ***p<0.001. Scale bars: 50 μm. Data are shown as the mean±SD (n=4 for each experiment mice group).