2019
DOI: 10.1080/20013078.2019.1697028
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Quality and efficiency assessment of six extracellular vesicle isolation methods by nano‐flow cytometry

Abstract: Extracellular vesicles (EVs) have sparked tremendous interest owing to their prominent potential in diagnostics and therapeutics. Isolation of EVs from complex biological fluids with high purity is essential to the accurate analysis of EV cargo. Unfortunately, generally used isolation techniques do not offer good separation of EVs from non-EV contaminants. Hence, it is important to have a standardized method to characterise the properties of EV preparations, including size distribution, particle concentration,… Show more

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Cited by 426 publications
(445 citation statements)
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References 64 publications
(97 reference statements)
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“…In this review, we mainly focused on the exosomes. However, because of absence of strict standards for exosome isolation and purification methods, the International Society for Extracellular Vesicles encouraged researchers to establish minimum requirements and strictly control the integrity, size, molecular cargo, and functionality of the vesicle population [38,[55][56][57], so that we narrowed the research of exosomes based on the widely accepted methods. Exosomes are small extracellular vesicles (40-160 nm in diameter) formed by dynamic exocytosis [58][59][60].…”
Section: The Biosynthesis and Function Of Extracellular Vesicles And mentioning
confidence: 99%
“…In this review, we mainly focused on the exosomes. However, because of absence of strict standards for exosome isolation and purification methods, the International Society for Extracellular Vesicles encouraged researchers to establish minimum requirements and strictly control the integrity, size, molecular cargo, and functionality of the vesicle population [38,[55][56][57], so that we narrowed the research of exosomes based on the widely accepted methods. Exosomes are small extracellular vesicles (40-160 nm in diameter) formed by dynamic exocytosis [58][59][60].…”
Section: The Biosynthesis and Function Of Extracellular Vesicles And mentioning
confidence: 99%
“…Looking at the literature, we noticed that the polymer kit consistently led to a greater number of vesicles detected by NanoSight [49,52,56], and yet led to a reduced detection of exosomal markers by Western blot [28,38,49,52,56,57]. Interestingly, Rider et al, while optimizing a polymer to extract extracellular vesicles, showed that rinsing of exosomes that had been precipitated using the polymer resulted in an increase in exosome markers detected by Western blot [28].…”
Section: Resultsmentioning
confidence: 96%
“…Several commercial kits have been developed to improve isolation efficacy and speed. The purity of vesicles isolated using these kits is often questioned in comparison to the ultracentrifugation methodology, especially when extracting from serum/plasma [49,50], but also in the in vitro context [51,52]. However, it is important to note that, whilst these studies do adhere strictly to the manufacturers' instructions for usage of the kits, they often fail to carry out identical sample preparations prior to the comparison -for example, carrying out centrifugations and/or filtration steps to remove microvesicles and other contaminants before ultracentrifugation but neglecting to do so before using the kits.…”
Section: Resultsmentioning
confidence: 99%
“…Several commercial kits have been developed to improve isolation e cacy and speed. The purity of vesicles isolated using these kits is often questioned in comparison to the ultracentrifugation methodology, especially when extracting from serum/plasma [56,57], but also in the in vitro context [58,59]. However, it is important to note that, whilst these studies do adhere strictly to the manufacturers' instructions for usage of the kits, they often fail to carry out identical sample preparations prior to the comparison -for example, carrying out centrifugations and/or ltration steps to remove microvesicles and other contaminants before ultracentrifugation but neglecting to do so before using the kits.…”
Section: Resultsmentioning
confidence: 99%