To evaluate the extent of tumor-associated lymphangiogenesis and angiogenesis, we quantified microvascular density of podoplanin+ and Lyve-1+ lymphatic vessels (LMVD) and of CD34+blood vessels (blood microvessel density)
α-SMA (CAF marker) and podoplanin (LEC marker，lymphatic vasculature)
Fibroblasts and CAFs expressed α-SMA, PDGFRβ, vimentin, and podoplanin, while fibroblasts and LECs expressed podoplanin, vimentin, VEGFR2 and VEGFR3
CAFs were isolated from fresh ICC tissues by the outgrowth method, as previously described.24 25
PMID: 34413131
结论：
GM-CSF neutralisation alone firmly deterred progression of autochthonous iCCA murine tumours and extended survival.
The LSL-KrasG12D; Trp53Flox/Flox; Alb-Cre (KPPC) mouse of spontaneous iCCA was previously described.14 15
CD68+ macrophages巨噬细胞 and CD15+ granulocytes粒细胞
trichrome staining (collage)，CD45 IHC staining (inflammatory leucocytes)
CD68 (TAM), CD15 (G-MDSC) and CD8 (cytotoxic T lymphocytes)
根据他人定义的免疫基因集对数据进行评分：
Four immune subtypes of human iCCA have been identified using 14 gene signatures derived from the MCP counter.20
Human microenvironment cell population (MCP) counter analysis was performed as previously described.17
Mouse MCP counter analysis was performed as previously described.18
