Interaction of rat tumor cells with blood vessels and lymphatics of the avian chorioallantoic membrane

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Abstract
It has generally been assumed that tumors do not induce lymphangiogenesis and only very recently animal models have been presented showing tumor‐induced lymphangiogenesis. We have grown two types of rat tumor cells, 10AS pancreatic carcinoma and C6 glioma cells, on the chorioallantoic membrane (CAM) of chick and quail embryos. The suspended tumor cells rapidly formed solid tumors which invaded the CAM and were vascularized by CAM vessels. When grown on the CAM of quail embryos intratumoral endothelial cells could be specifically stained with the QH1 antibody. In C6 gliomas the vascular pattern was more regular than in 10AS carcinomas. The vessels often grew radially into the glioma and many of them were invested by smooth muscle α‐actin‐positive periendothelial cells. Lymphatics, which were identified by vascular endothelial growth factor receptor‐3 (VEGFR‐3) in situ hybridization were absent from C6 gliomas, although a weak expression of the lymphangiogenic growth factor, VEGF‐C, could be detected in the C6 cells by Northern blot analysis. In contrast, 10AS cells, which expressed high levels of VEGF‐C, induced ingrowth of lymphatics into the tumors, with BrdU‐labeling rates of about 9% of lymphatic endothelial cells. Our studies demonstrate the heterogeneity of interactions of tumor cells with blood vessels and lymphatics and show that sufficient quantities and/or quality of lymphangiogenic growth factors are crucial for the induction of lymphatics in tumors. Microsc. Res. Tech. 55:100–107, 2001.

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