The Study On The Anti-Tongue Cancer Effects Induced By Exosome Pulsed Dendritic Cells

Part one:Isolation and analysis of tumor-derived exosomesObjective To investigate whether the tumor cell line Tca-8113 could secrete exosomes when culture in vitro; and analyze the exosomes in their size, shape and composition.Material and Methods Tca-8113 cells were cultured with RPIM1640, isolated and purified the tumor-derived exosomes from the culture supernatants by ultrafiltration with Millipore Centrifual Filter Devices; analyzed by Electron Microscopy, Western blotting, and ELISA. In addition, frozen and thawed Tca-8113 cells to get frozen tumor antigens (FTA).Results Electron microscopy analysis of Tca-8113 squamous carcinoma cell line revealed numerous MVBs in the cells. These compartments contained internal vesicles that were released into extracellular medium when the membrane of MVB fused with plasma membrane. In the culture supernatants of Tca-8113 cells, the vesicles—exosomes can be found, with ball, ellipse and dumbbell shapes, and 30 - 90 nm in diameter, often aggregated. Quantitative analyses of exosomes showed that a mean of 7.39× 10~9 exosomal MHC class I molecules can be harvested from 1 ml supernatant; MHC class I molecule and Hsc70 in exosomes and FTA were positive when analyzed by Western blotting.