Open Access

Scientific Reports, volume 5, issue 1, publication number 17319

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

Livshits Mikhail A ^{1, 2}

Khomyakova Elena ³

Evtushenko Evgeniy G ⁴

Lazarev Vassili N. ^{2, 3}

Kulemin Nikolay A. ^{2, 3}

Semina Svetlana E ⁵

Generozov Edward V. ³

GOVORUN VADIM M. ³

Hide authors affiliations Show authors affiliations: 5 affiliations

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia |

Moscow Institute of Physics and Technology, Dolgoprudny, Russia |

Fededal Research and Clinical Center of Physical-Chemical Medicine, Moscow, Russia |

⁴

Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia |

⁵

Research Institute of Carcinogenesis, N. N. Blokhin Russian Cancer Research Center, Moscow, Russia |

Publication type: Journal Article

Publication date: 2015-11-30

Springer Nature

Journal: Scientific Reports

Quartile SCImago

Quartile WOS

Impact factor: 4.6

ISSN: 20452322

DOI: 10.1038/srep17319

Copy DOI

PubMed ID: 26616523

Multidisciplinary

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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	10 20 30 40 50 60 70 80 90

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Livshits M. A. et al. Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol // Scientific Reports. 2015. Vol. 5. No. 1. 17319

GOST all authors (up to 50) Copy

Livshits M. A., Khomyakova E., Evtushenko E. G., Lazarev V. N., Kulemin N. A., Semina S. E., Generozov E. V., GOVORUN V. M. Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol // Scientific Reports. 2015. Vol. 5. No. 1. 17319

RIS |

Cite this

RIS Copy

TY - JOUR

DO - 10.1038/srep17319

UR - https://doi.org/10.1038%2Fsrep17319

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

T2 - Scientific Reports

AU - Livshits, Mikhail A

AU - Khomyakova, Elena

AU - Evtushenko, Evgeniy G

AU - Lazarev, Vassili N.

AU - Kulemin, Nikolay A.

AU - Semina, Svetlana E

AU - Generozov, Edward V.

AU - GOVORUN, VADIM M.

PY - 2015

DA - 2015/11/30 00:00:00

PB - Springer Nature

IS - 1

VL - 5

PMID - 26616523

SN - 2045-2322

ER -

BibTex

Cite this

BibTex Copy

@article{2015_Livshits,

author = {Mikhail A Livshits and Elena Khomyakova and Evgeniy G Evtushenko and Vassili N. Lazarev and Nikolay A. Kulemin and Svetlana E Semina and Edward V. Generozov and VADIM M. GOVORUN},

title = {Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol},

journal = {Scientific Reports},

year = {2015},

volume = {5},

publisher = {Springer Nature},

month = {nov},

url = {https://doi.org/10.1038%2Fsrep17319},

number = {1},

doi = {10.1038/srep17319}

}

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Springer Nature

Journal

Scientific Reports

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Quartile WOS

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4.6

ISSN

20452322 (Print)

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Laboratory of Human Molecular Genetics

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