Transmission electron microscopy assessment of a novel method for isolating pure exosomes from serum

被引:4
作者
Wen, Jin [1 ]
Zhang, Zhijun [1 ]
Feng, Gang [2 ]
Zhang, Yang [3 ]
Li, Hanzhong [1 ,7 ]
Lambert, Claude [4 ]
Mallouk, Nora [5 ]
Li, Guorong [6 ]
机构
[1] Peking Union Med Coll Hosp, Dept Urol, Beijing, Peoples R China
[2] Wannan Med Coll, Clin Lab, Affiliated Hosp 1, Wuhu, Peoples R China
[3] Guangzhou HopeTech Biol Technol Co LTD, Guangzhou, Peoples R China
[4] CHU St Etienne, North Hosp, Lab Flow Cytometry, Immunol Lab, St Etienne, France
[5] Univ Jean Monnet, Fac Med, Ctr Elect Microscopy, CMES, St Etienne, France
[6] CHU St Etienne, North Hosp, Dept Urol, St Etienne, France
[7] Beijing Chinese Acad Med Sci & Peking Union Med Co, Peking Union Med Coll Hosp, Dept Urol, Beijing 100730, Peoples R China
关键词
Electron microscopy; exosomes; flow cytometry; human; isolation; serum;
D O I
10.1080/10520295.2023.2202415
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Serum exosomes frequently are used for liquid biopsy. Serum exosomes normally are isolated using ultracentrifugation; however, ultracentrifugation is time-consuming, labor intensive and requires a high-speed centrifuge. Many commercial kits use a precipitation-based method; however, this process can result in substantial contamination. We developed a new method to isolate pure serum exosomes. We isolated serum exosomes using precipitation, extracted them using acetone, then isolated them again by precipitation. We used transmission electron microscopy (TEM) to examine the morphology of serum exosomes. TEM indicated that our isolated exosomes were pure with typical morphology and with a size ranging from 40 to 150 nm. Flow cytometry revealed expression of exosome markers, CD63, CA81 and CD9. Our double precipitation method enables ready extraction of pure exosomes from serum. Our double precipitation method simplifies detection of serum exosomal biomarkers for diagnosis and prognosis of disease.
引用
收藏
页码:391 / 395
页数:5
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