Isolation, Characterization, and Cytotoxicity Investigation of the Exosomes Secreted by Doxorubicin Primed Cells

Abstract

Introduction :Exosomes are natural nanosized carriers that have many advantages, including non-toxic and non-immunogenic, and they have the ability to be designed for specific targeting and drug delivery, which makes them useful as a powerful nanostructure to deliver anti-cancer drugs with the aim of cancer treatment. Aim :Isolation, characterization, and cytotoxicity investigation of the exosomes secreted by doxorubicin primed cells. Methods: In this study, exosomes were isolated from the cell culture media containing doxorubicin using a commercial kit available in the market, and in terms of physico-chemical characteristics; Surface markers, surface charge, particle size, and shape will be examined by TEM and SEM microscope and DLS, and Western blot and BCA protein test and studing on cytotoxicity effect by MTT test and investigating drug penetration inside the cells. Results: The extracted exosomes were studied by TEM/SEM and DLS. The average size of exosomes obtained by DLS was 100-120 nm. The average size of exosomes calculated by SEM is 82-104 nm. Western blot analysis also showed the expression of (CD81, CD63) specific markers of exosome in the purified exosomes. Also, BCA test showed linear graph and average protein amount calculated 2269.035ug/ml . the cytotoxicity of the isolated exosomes will be evaluated using the MTT test.in MTT test by increasing drug concentration toxic effects increased but it didnot show significant effect on exosomes that secreted from non-treated and treated cells with doxorubicin drug. Investigating drug uptake showed that drug can penetrated inside the cells.Conclusion: Exosomes that extracted from media containing Doxorubicin didinot show significant effect on tumor cells .