Separation and Identification of the Exosomes Derived from A Human Tumor Cell Line
10.3969/j.issn.0253-9896.2009.12.007
- VernacularTitle:肿瘤细胞来源胞外体的分离鉴定与功能检测
- Author:
Yan CUI
;
Jinpu YU
;
Hui LI
;
Lili YANG
;
Feng WEI
;
Xiumei AN
;
Xiubao REN
- Publication Type:Journal Article
- Keywords:
cell line;
tumor breast neoplasms immune tolerance cell separation tumor cells;
cultured
- From:
Tianjin Medical Journal
2009;37(12):1014-1016,彩1
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To separate and identify the exosomes derived from MCF-7 tumor cells,and observe their function on IL-2 induced lymphocyte proliferation.Methods:The exosomes were separated by serial centrifugation,concentration and density gradient centrifugation, and then identified by transmission electron microscope.The effects of exosomes on IL-2 induced proliferation were studied by MTT method.Results:Total 20 mL exosomes from 2×10~7 MCF-7 cells with concentration of protein were(1.84±0.01) g/L. The diameters of these purified bi-layer membrane exosomes were 55-70 nm, which were consistent with the internal vesicles in multi-vesicular endosomes.The result of MTT test showed that proliferation of donor peripheral blood lymphocytes was inhibited by tumor exosomes at a dose-dependent manner in response to IL-2.Conclusion:It is feasible to separate high-purity exosomes from tumor cell culture medium supernatant by serial centrifugation and concentrate method. These tumor derived exosomes have inhibitory effect on IL-2 induced proliferation.
