Open AccessIsolating Exosomes Derived From Human Natural Killer Cells For Characterization
Author(s) -
Wesley Huang,
Jo Carmona
Publication year - 2018
Publication title -
journal of student research
Language(s) - English
Resource type - Journals
ISSN - 2167-1907
DOI - 10.47611/jsrhs.v7i2.471
Subject(s) - microvesicles , cd81 , cytotoxic t cell , granzyme b , cytotoxicity , exosome , biology , microbiology and biotechnology , chemistry , luciferase , microrna , cell culture , immunology , in vitro , biochemistry , transfection , genetics , gene , hepatitis c virus , virus
Exosomes have been an area of great interest in the field of research, as they seem to participate in many essential cellular processes. However, current methods in isolating exosomes have many limitations. The aim of this study is to present a more effective, efficient, and simple method to isolate large quantities of natural killer (NK) cell-derived exosomes using the ÄKTA start system, centered around size-exclusion chromatography, and to characterize these exosomes. Using the ÄKTA system, we obtained a chromatogram of high resolution depicting the UV absorbance of exosomes. We confirmed the identity of the exosomes by probing for markers such as CD56 and CD81 and cytotoxic proteins such as granzyme B in the exosomes, whose cytotoxic properties against cancer cells were confirmed using luciferase assays. By incubating exosomes, cancer cells, and an array of inhibitors, we also found that NK exosomes may engage in cytotoxicity via different methods such as ligand-receptor interactions.