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

Sidhom, Karim; Patience, O.; Saleem, Ayesha

doi:10.20944/preprints202007.0485.v1

Cited by 21 publications

(30 citation statements)

References 57 publications

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“…These particles often produce significant challenges for downstream application/analysis. 31 Figures 1G and 1I. Therefore, the SEM based approach can offer several benefits over the other approaches as the morphology and size can be monitored concomitantly.…”

Section: Discussionmentioning

confidence: 99%

High throughput imaging of nanoscale extracellular vesicles by scanning electron microscopy for accurate size-based profiling and morphological analysis

Cavallaro

Hååg

Viktorsson

et al. 2021

Preprint

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Nanoscale extracellular vesicle (EVs) have been found to play a key role in intercellular communication, offering opportunities for both diagnostics and therapeutics. However, lying below the diffraction limit and also being highly heterogeneous in their size, morphology and abundance, these vesicles pose significant challenges for their physical characterization. Here, we present a direct visual approach for their accurate morphological and size-based profiling by using scanning electron microscopy (SEM). To achieve that, we methodically examined various process steps and developed a protocol to improve the throughput, conformity and image quality while preserving the shape of EVs. The investigation was performed with small EVs (sEVs) isolated from a non-small cell lung cancer (NSCLC) cell line H1975 as well as from a human serum, and the results were compared with those obtained from nanoparticle tracking analysis (NTA). While the comparison of the sEV size distributions showed good agreement between the two methods for large sEVs (diameter >70 nm), the microscopy based approach showed a better capacity for analyses on smaller vesicles, with higher sEV counts compared to NTA. In addition, we demonstrated the possibility of identifying non-EV particles based on size and morphological features. The study also showed process steps that can generate artifacts bearing resemblance with sEVs. The results therefore present a simple way to use a widely available microscopy tool for accurate and high throughput physical characterization of EVs.

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Section: Discussionmentioning

confidence: 99%

High throughput imaging of nanoscale extracellular vesicles by scanning electron microscopy for accurate size-based profiling and morphological analysis

Cavallaro

Hååg

Viktorsson

et al. 2021

Preprint

View full text Add to dashboard Cite

show abstract

“…The lower membrane captures EVs < 200 nm or >20 nm, while larger vesicles are retained on the upper filter, and the smallest vesicles are discarded [ 100 ]. Size-exclusion chromatography also isolates EVs by size: smaller molecules slow down because they enter the pores of the gel, while EVs do not enter the pores and flow through faster [ 101 ] ( Figure 4(d) ). This technique can preserve EV structure better than ultracentrifugation and ultrafiltration.…”

Section: Isolation Of Evsmentioning

confidence: 99%

Extracellular Vesicles in Liquid Biopsies: Potential for Disease Diagnosis

Liu

Chen

Pei

et al. 2021

BioMed Research International

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Liquid biopsy is conducted through minimally invasive or noninvasive procedures, and the resulting material can be subjected to genomic, proteomic, and lipidomic analyses for early diagnosis of cancers and other diseases. Extracellular vesicles (EVs), one kind of promising tool for liquid biopsy, are nanosized bilayer particles that are secreted by all kinds of cells and that carry cargoes such as lipids, proteins, and nucleic acids, protecting them from enzymatic degradation in the extracellular environment. In this review, we provide a comprehensive introduction to the properties and applications of EVs, including their biogenesis, contents, sample collection, isolation, and applications in diagnostics based on liquid biopsy.

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“…Therefore, the large-scale production of exosomes might be expensive [ 118 ]. However, it should be kept in mind that recent developments in the field of exosome isolation, such as size exclusion chromatography, may significantly improve scalability and costs in the near future [ 131 ].…”

Section: Non-cellular and Bacterial Agents For Igsmentioning

confidence: 99%

Cell-Based Tracers as Trojan Horses for Image-Guided Surgery

Sier

Vries

Vorst

et al. 2021

IJMS

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Surgeons rely almost completely on their own vision and palpation to recognize affected tissues during surgery. Consequently, they are often unable to distinguish between different cells and tissue types. This makes accurate and complete resection cumbersome. Targeted image-guided surgery (IGS) provides a solution by enabling real-time tissue recognition. Most current targeting agents (tracers) consist of antibodies or peptides equipped with a radiolabel for Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT), magnetic resonance imaging (MRI) labels, or a near-infrared fluorescent (NIRF) dye. These tracers are preoperatively administered to patients, home in on targeted cells or tissues, and are visualized in the operating room via dedicated imaging systems. Instead of using these ‘passive’ tracers, there are other, more ‘active’ approaches of probe delivery conceivable by using living cells (macrophages/monocytes, neutrophils, T cells, mesenchymal stromal cells), cell(-derived) fragments (platelets, extracellular vesicles (exosomes)), and microorganisms (bacteria, viruses) or, alternatively, ‘humanized’ nanoparticles. Compared with current tracers, these active contrast agents might be more efficient for the specific targeting of tumors or other pathological tissues (e.g., atherosclerotic plaques). This review provides an overview of the arsenal of possibilities applicable for the concept of cell-based tracers for IGS.

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

Cited by 21 publications

References 57 publications

High throughput imaging of nanoscale extracellular vesicles by scanning electron microscopy for accurate size-based profiling and morphological analysis

High throughput imaging of nanoscale extracellular vesicles by scanning electron microscopy for accurate size-based profiling and morphological analysis

Extracellular Vesicles in Liquid Biopsies: Potential for Disease Diagnosis

Cell-Based Tracers as Trojan Horses for Image-Guided Surgery

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