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

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,...

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Vydáno v:International journal of molecular sciences Ročník 21; číslo 18; s. 6466
Hlavní autoři: Sidhom, Karim, Obi, Patience O., Saleem, Ayesha
Médium: Journal Article
Jazyk:angličtina
Vydáno: Switzerland MDPI AG 04.09.2020
MDPI
Témata:
Animals
Chromatography
Chromatography, Gel - methods
Communication
Deoxyribonucleic acid
DNA
Exosomes - chemistry
Exosomes - metabolism
Extracellular vesicles
Extracellular Vesicles - chemistry
Extracellular Vesicles - metabolism
Humans
Membranes
MicroRNAs
Mitochondrial DNA
Pore size
Proteins
Review
Sedimentation & deposition
Ultracentrifugation - methods
Ultrafiltration - methods
extracellular vesicles
exosomes
size-exclusion chromatography
immunoaffinity capture
poly-ethylene glycol
microvesicles
differential ultracentrifugation
microfluidics
ISSN:1422-0067, 1661-6596, 1422-0067
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Shrnutí: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.
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These authors contributed equally to this work.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms21186466