Comparison of Plasma Exosomes by Differential Ultracentrifugation and Solvent Precipitation Methods

被引：12
作者：
Peng, Qiao [1 ,2 ]
Zhang, Jing [1 ,2 ,3 ]
Zhou, Gang [1 ,2 ,3 ]
机构：
[1] Wuhan Univ, Sch & Hosp Stomatol, State Key Lab Breeding Base Basic Sci Stomatol Hu, Wuhan, Hubei, Peoples R China
[2] Wuhan Univ, Sch & Hosp Stomatol, Minist Educ, Key Lab Oral Biomed, Wuhan, Hubei, Peoples R China
[3] Wuhan Univ, Sch & Hosp Stomatol, Dept Oral Med, Luoyu Rd 237, Wuhan, Hubei, Peoples R China
来源：
CLINICAL LABORATORY | 2018年 / 64卷 / 06期
基金：
中国国家自然科学基金;
关键词：
exosomes; oral lichen planus; plasma; differential ultra-centrifugation; ORAL LICHEN-PLANUS; T-CELLS; BIOMARKERS; AUTOPHAGY; DISEASE; EXPRESSION; MICRORNA; THERAPY; ROLES;
D O I：
10.7754/Clin.Lab.2018.180104
中图分类号：
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号：
1001 ;
摘要：
Background: Emerging evidence has identified that exosomes play a pivotal role in intercellular signal transmission. However, the standardized purification techniques to isolate high quality exosomes are still deficient at present. This study was to evaluate reproducibility and efficiency of differential ultracentrifugation and solvent precipitation-based kits by isolating plasma-derived exosomes from oral lichen planus patients. Methods: Morphology, exosomal biomarkers, particle size distribution, proteomic components, and protein yield of isolated exosomes were evaluated by transmission electron microscope, western blot, laser diffraction instrument, Coomassie staining, and BCA protein assay kit, respectively. Results: TEM displayed representative cup-shaped morphology of exosomes and western blot identified exosomal biomarkers CD9 and CD63. The size distribution showed that particles by differential ultracentrifugation were mainly from 26.15 nm to 166.5 nm, while some of the particles obtained by solvent precipitation kits were larger than 1,000 nm. In addition, exosomes isolated by solvent precipitation kits showed a significantly higher amount of protein yield due to plasma albumin contamination. Conclusions: Both differential ultracentrifugation and precipitation based kits could successfully isolate plasma exosomes, and exosomes by differential ultracentrifugation were purer and more appropriate for further proteomic analysis.
引用
收藏
页码：991 / 998
页数：8
相关论文
共 31 条
[11]   Exosomes: Vehicles of Intercellular Signaling, Biomarkers, and Vectors of Cell Therapy [J].
Kourembanas, Stella .
ANNUAL REVIEW OF PHYSIOLOGY, VOL 77, 2015, 77 :13-27
[12]   Human aortic smooth muscle cell-derived exosomal miR-221/222 inhibits autophagy via a PTEN/Akt signaling pathway in human umbilical vein endothelial cells [J].
Li, Luocheng ;
Wang, Zhiwei ;
Hu, Xiaoping ;
Wan, Ting ;
Wu, Hongbing ;
Jiang, Wanli ;
Hu, Rui .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2016, 479 (02) :343-350
[13]   Progress in Exosome Isolation Techniques [J].
Li, Pin ;
Kaslan, Melisa ;
Lee, Sze Han ;
Yao, Justin ;
Gao, Zhiqiang .
THERANOSTICS, 2017, 7 (03) :789-804
[14]   Exosomes Derived from Mesenchymal Stem Cells Rescue Myocardial Ischaemia/Reperfusion Injury by Inducing Cardiomyocyte Autophagy Via AMPK and Akt Pathways [J].
Liu, Liang ;
Jin, Xian ;
Hu, Cui-Fen ;
Li, Rong ;
Zhou, Zhong'e ;
Shen, Cheng-Xing .
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, 2017, 43 (01) :52-68
[15]   Exosomes released by keratinocytes modulate melanocyte pigmentation [J].
Lo Cicero, Alessandra ;
Delevoye, Cedric ;
Gilles-Marsens, Floriane ;
Loew, Damarys ;
Dingli, Florent ;
Guere, Christelle ;
Andre, Nathalie ;
Vie, Katell ;
van Niel, Guillaume ;
Raposo, Graca .
NATURE COMMUNICATIONS, 2015, 6
[16]   Optimized exosome isolation protocol for cell culture supernatant and human plasma [J].
Lobb, Richard J. ;
Becker, Melanie ;
Wen, Shu Wen ;
Wong, Christina S. F. ;
Wiegmans, Adrian P. ;
Leimgruber, Antoine ;
Moeller, Andreas .
JOURNAL OF EXTRACELLULAR VESICLES, 2015, 4
[17]   Oral candidiasis following steroid therapy for oral lichen planus [J].
Marable, D. R. ;
Bowers, L. M. ;
Stout, T. L. ;
Stewart, C. M. ;
Berg, K. M. ;
Sankar, V. ;
DeRossi, S. S. ;
Thoppay, J. R. ;
Brennan, M. T. .
ORAL DISEASES, 2016, 22 (02) :140-147
[18]  
Mincheva-Nilsson L, 2016, CURR PROTOC IMMUNOL, V115
[19]   The multifaceted exosome: Biogenesis, role in normal and aberrant cellular function, and frontiers for pharmacological and biomarker opportunities [J].
Pant, Saumya ;
Hilton, Holly ;
Burczynski, Michael E. .
BIOCHEMICAL PHARMACOLOGY, 2012, 83 (11) :1484-1494
[20]   Autophagy and the endo/exosomal pathways in health and disease [J].
Papandreou, Margarita-Elena ;
Tavernarakis, Nektarios .
BIOTECHNOLOGY JOURNAL, 2017, 12 (01)
1234