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Cited by 686

The DOI matches its Crossref record; 33 of the 33 references it cites were found in a registry or an index.

Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol

Open access
Livshits, Mikhail A., Khomyakova, Elena, Evtushenko, Evgeniy G., Lazarev, Vassili N., Kulemin, Nikolay A., Semina, Svetlana E., Generozov, Edward V., Govorun, Vadim M.·2015·Scientific Reports
DOI 10.1038/srep17319
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Abstract

Exosomes, small (40–100 nm) extracellular membranous vesicles, attract enormous research interest because they are carriers of disease markers and a prospective delivery system for therapeutic agents. Differential centrifugation, the prevalent method of exosome isolation, frequently produces dissimilar and improper results because of the faulty practice of using a common centrifugation protocol with different rotors. Moreover, as recommended by suppliers, adjusting the centrifugation duration according to rotor K-factors does not work for “fixed-angle” rotors. For both types of rotors – “swinging bucket” and “fixed-angle” – we express the theoretically expected proportion of pelleted vesicles of a given size and the “cut-off” size of completely sedimented vesicles as dependent on the centrifugation force and duration and the sedimentation path-lengths. The proper centrifugation conditions can be selected using relatively simple theoretical estimates of the “cut-off” sizes of vesicles. Experimental verification on exosomes isolated from HT29 cell culture supernatant confirmed the main theoretical statements. Measured by the nanoparticle tracking analysis (NTA) technique, the concentration and size distribution of the vesicles after centrifugation agree with those theoretically expected. To simplify this “cut-off”-size-based adjustment of centrifugation protocol for any rotor, we developed a web-calculator.

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Crossref · 10.1038/srep17319

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Metadata

Authors
Livshits, M. A., Khomyakova, E., Evtushenko, E. G., Lazarev, V. N., Kulemin, N. A., Semina, S. E., Generozov, E. V., & Govorun, V. M.
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2015-11-30
Volume
5
Issue
1
Pages
17319-17319
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/srep17319
OpenAlex
W2414083225
Cited by
686
References
33
FWCI
9.39
Type
journal article
Language
en
Open access
gold

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