THP-1分化的巨噬细胞来源Exosomes的生物学特征鉴定  

Analysis of the Biological Characterization of Exosomes Secreted from THP-1 Differentiation Macrophage

作　　者：王建军[1] 姚永良[1] 陈偲[2] 吴建红[1] 赵淑玲[2] 汤立军[2] 

机构地区：[1]江苏大学附属昆山医院,中国江苏昆山215300 [2]中南大学生命科学学院分子生物学系,中国湖南长沙410078

出　　处：《生命科学研究》2014年第4期283-285,309,共4页Life Science Research

基　　金：国家自然科学基金资助项目(81071326);昆山市科技计划项目(KS1347)

摘　　要：利用佛波酯体外分化人急性单核白血病细胞形成巨噬细胞,并用低温超速离心法从巨噬细胞培养上清收集Exosomes,并分析其生物学特征。Exosomes经透射电显微镜分析形态,并用Western blot方法分析Exosomes膜上特异性蛋白CD80、ICAM-3、HSP-70的表达对其生物学特征进行鉴定。研究结果表明佛波酯成功诱导人急性单核白血病细胞分化成人巨噬细胞。获得的巨噬细胞分泌微小囊泡经透射电镜分析后,微小囊泡的直径在30-100 nm,并且其含有Exosomes的特征性蛋白CD80、ICAM-3、HSP-70等蛋白,表明其为Exosomes。使用低温超速离心法成功收集到Exosomes,为后续开展Exosomes的成份及功能分析提供了保障。Human acute monocytic leukemia cells were induced to differentiate into macrophages with phorbol myristoyl acetate（PMA） in vitro, then Exosomes were purified from the culture medium of macrophages supernatant by ultracentrifugation method. The morphology of Exosomes were analyzed by transmission electron microscopy, and specific membrane proteins such as CD80, ICAM-3, and HSP-70 were detected by Western blot. The results show that human acute monocytic leukemia cells were induced into human macrophages successfully. The diameter of tiny vesicles analyzed by transmission electron microscopy（TEM）was 30-100 nm, and Exosomes specific proteins CD80, ICAM-3, and HSP-70 were also be detected successfully. Above all mean Exosomes could be purified by the ultracentrifugation method effectively, and laid a solid functional and components analysis of Exosomes in the following experiments.

关 键 词：巨噬细胞 EXOSOMES 透射电镜 超速离心 

分 类 号：Q28[生物学—细胞生物学]