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Title: Effect of exosome isolation methods on physicochemical properties of exosomes and clearance of exosomes from the blood circulation
Authors: Yamashita, Takuma
Takahashi, Yuki  kyouindb  KAKEN_id
Nishikawa, Makiya
Takakura, Yoshinobu  kyouindb  KAKEN_id
Author's alias: 高橋, 有己
高倉, 喜信
Keywords: Ultracentrifugation
Tunable resistive pulse sensing
Sterile filtration
Density gradient centrifugation
Issue Date: Jan-2016
Publisher: Elsevier B.V.
Journal title: European Journal of Pharmaceutics and Biopharmaceutics
Volume: 98
Start page: 1
End page: 8
Abstract: Exosomes, which are expected to be delivery systems for biomolecules such as nucleic acids, are collected by several methods. However, the effect of exosome isolation methods on the characteristics of exosomes as drug carriers, such as recovery efficiency after sterile filtration and pharmacokinetics, has not been investigated despite the importance of these characteristics for the development of exosome-based delivery systems. In the present study, exosomes collected from murine melanoma B16-BL6 cells by several methods were compared with respect to dispersibility, recovery rate after filtering, and clearance from the blood circulation in mice. The exosomes were collected by three ultracentrifugation-based methods: simple ultracentrifugation/pelleting (pelleting method), ultracentrifugation with an iodixanol cushion (cushion method), and ultracentrifugation on an iodixanol density gradient (gradient method). The isolation methods had little effect on the particle number of exosomes. In contrast, transmission electron microscopy observation and size distribution measurement using tunable resistive pulse sensing indicated that the exosomes of the gradient method were more dispersed than the others. The exosomes were labeled with Gaussia luciferase and intravenously injected into mice. Clearance of injected exosomes from the blood circulation did not significantly change with isolation methods. When the exosomes were filtered using a 0.2-μm filter, the recovery rate was 82% for the exosomes of the gradient method, whereas it was less than 50% for the others. These results indicate that the exosome isolation method markedly affects the dispersibility and filtration efficiency of the exosomes.
Rights: © 2015. This manuscript version is made available under the CC-BY-NC-ND 4.0 license
The full-text file will be made open to the public on 1 January 2017 in accordance with publisher's 'Terms and Conditions for Self-Archiving'
This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。
DOI(Published Version): 10.1016/j.ejpb.2015.10.017
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