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Open AccessReview

A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option?

by Karim Sidhom 1,2,3,4,†, Patience O. Obi 2,3,4,5,6,† and Ayesha Saleem 2,3,4,5,6,*
1
Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB R3E 3P5, Canada
2
Diabetes Research Envisioned and Accomplished in Manitoba (DREAM) Research Theme of CHRIM, Winnipeg, MB R3E 3P4, Canada
3
Biology of Breathing Research Theme of CHRIM, Winnipeg, MB R3E 3P4, Canada
4
Children’s Hospital Research Institute of Manitoba (CHRIM), Winnipeg, MB R3E 3P4, Canada
5
Applied Health Sciences, Faculty of Graduate Studies, University of Manitoba, Winnipeg, MB R3T 2N2, Canada
6
Faculty of Kinesiology and Recreation Management, University of Manitoba, Winnipeg, MB R3T 2N2, Canada
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(18), 6466; https://doi.org/10.3390/ijms21186466
Received: 20 July 2020 / Revised: 29 August 2020 / Accepted: 31 August 2020 / Published: 4 September 2020
(This article belongs to the Special Issue Role of the Extracellular Vesicles in Obesity and Related Diseases)
Extracellular vesicles (EVs) are membranous vesicles secreted by both prokaryotic and eukaryotic cells and play a vital role in intercellular communication. EVs are classified into several subtypes based on their origin, physical characteristics, and biomolecular makeup. Exosomes, a subtype of EVs, are released by the fusion of multivesicular bodies (MVB) with the plasma membrane of the cell. Several methods have been described in literature to isolate exosomes from biofluids including blood, urine, milk, and cell culture media, among others. While differential ultracentrifugation (dUC) has been widely used to isolate exosomes, other techniques including ultrafiltration, precipitating agents such as poly-ethylene glycol (PEG), immunoaffinity capture, microfluidics, and size-exclusion chromatography (SEC) have emerged as credible alternatives with pros and cons associated with each. In this review, we provide a summary of commonly used exosomal isolation techniques with a focus on SEC as an ideal methodology. We evaluate the efficacy of SEC to isolate exosomes from an array of biological fluids, with a particular focus on its application to adipose tissue-derived exosomes. We argue that exosomes isolated via SEC are relatively pure and functional, and that this methodology is reproducible, scalable, inexpensive, and does not require specialized equipment or user expertise. However, it must be noted that while SEC is a good candidate method to isolate exosomes, direct comparative studies are required to support this conclusion. View Full-Text
Keywords: extracellular vesicles; exosomes; microvesicles; differential ultracentrifugation; poly-ethylene glycol; immunoaffinity capture; microfluidics; size-exclusion chromatography extracellular vesicles; exosomes; microvesicles; differential ultracentrifugation; poly-ethylene glycol; immunoaffinity capture; microfluidics; size-exclusion chromatography
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Sidhom, K.; Obi, P.O.; Saleem, A. A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option? Int. J. Mol. Sci. 2020, 21, 6466.

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