Effect of pancreatic cancer derived extracellular vesicles on bone marrow drived macrophages

Our results show that PaCa cell derived EVs were taken up by the macrophage in a time-dependent manner that resulted in an immunosuppressive phenotype characterized by higher expression of CD206 and PD-L1 and higher secretion of immunosuppressive cytokines including TGF-beta, IL-10 and GM-CSF. Overall design: EVs were isolated from the conditioned medium that was used to culture one established pancreatic cancer cell (PANC-1), one PDX cell line (PPCL68) and a normal (non-tumorigenic) established pancreatic epithelial cell line (hTERT-HPNE), using size exclusion chromatography. Macrophages were generated from the bone marrow cells of C57/BL6 mice cultured in vitro using macrophage colony stimulating factor (M-CSF). The macrophages were seeded into 24 well plates and co-cultured with isolated PaCa or normal cell derived EVs at the concentration of 10 ug/mL EVs for 48h.