临床神经外科杂志2017，Vol.14Issue(5)：321-325,5.DOI:10.3969/j.issn.1672-7770.2017.05.001
胶质瘤细胞来源外泌体的提取与鉴定
Isolation and identification of exosomes originated from human glioma cells
摘要
Abstract
Objective To establish a method to extracte and identify exosomes from human glioma cells for further study .Methods Exosomes in the culture media of U 251 glioma cells were isolated by ultracentrifugation,and then characterized by transmission electron microscope (TEM), nanoparticle size analyzer ,and finally identified the exosome molecular markers CD 9 and CD63 by immunoblotting.Results Examined by TEM,the extraction of U251 glioma cell culture media has cupped microvesicles surrounded by an intact double membrane .The diameters of microvesicles approximately ranged from 30 to 150 nm.The results of nanoparticle size analyzer measurement showed that the diameters of microvesicles were about 50~150 nm,similar to the results by TEM . Western blotting analysis exhibited that the microvesicles express CD 9 and CD63 , the molecular markers for exosomes .Conclusion The microvesicles extracted from U 251 cell culture media by ultracentrifugation are exosomes and could be used for further study .In addition,a mature and stable method for exosome isolation has been established .关键词
人胶质瘤细胞/外泌体/分离/鉴定Key words
human glioma cell/exosome/isolation/identification分类
医药卫生引用本文复制引用
王维冰,吴世双,付佳乐,于如同,周秀萍..胶质瘤细胞来源外泌体的提取与鉴定[J].临床神经外科杂志,2017,14(5):321-325,5.基金项目
国家自然科学基金(81372699,81472345,81672489) （81372699,81472345,81672489）
江苏省"333"工程(BRA2015394) （BRA2015394）
江苏省"六大人才高峰"(WSW-039) （WSW-039）
