Assessment of separation methods for extracellular vesicles from human and mouse brain tissues and human cerebrospinal fluids

Muraoka, Satoshi; Lin, Weiwei; Chen, Mei; Hersh, Samuel; Emili, Andrew; Xia, Weiming; Ikezu, Tsuneya

doi:10.1016/j.ymeth.2020.02.002

Cited by 49 publications

(79 citation statements)

References 64 publications

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“…Last but not least, flow cytometry (FC), commonly used for the analysis of cells, is being actively developed for EV analysis [146,147] and is adopted by an increasing number of research groups, mainly to study the larger EVs [148,149]. Direct FC analysis using tracers that stain the whole vital EV compartment, such as lipophilic carbocyanine dyes, combined with phalloidin, that selectively binds to F-actin, accurately discriminates EVs from artifacts [42][43][44]150]. Standardized FC has a high potential for the detection of EVs in body fluids and, when combined with specific antibodies, concurrently allows EV immunophenotypic characterization [42,44,84,131,151,152] ( Figure 2).…”

Section: Methodological Approachesmentioning

confidence: 99%

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Simeone

Bologna

Lanuti

et al. 2020

IJMS

132

117

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Extracellular vesicles act as shuttle vectors or signal transducers that can deliver specific biological information and have progressively emerged as key regulators of organized communities of cells within multicellular organisms in health and disease. Here, we survey the evolutionary origin, general characteristics, and biological significance of extracellular vesicles as mediators of intercellular signaling, discuss the various subtypes of extracellular vesicles thus far described and the principal methodological approaches to their study, and review the role of extracellular vesicles in tumorigenesis, immunity, non-synaptic neural communication, vascular-neural communication through the blood-brain barrier, renal pathophysiology, and embryo-fetal/maternal communication through the placenta.

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Section: Methodological Approachesmentioning

confidence: 99%

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Simeone

Bologna

Lanuti

et al. 2020

IJMS

132

117

View full text Add to dashboard Cite

show abstract

“…EV pellets are often contaminated by other high abundant molecules (e.g., lipoproteins, protein aggregates, soluble proteins) or proteins that bind non-specifically to the exosomes and can interfere with further MS analysis. A density gradient flotation, such as the sucrose gradient [ 86 ] or the iodixanol (OptiPrepTM) velocity gradient [ 87 , 88 ], can be applied to the differential ultracentrifugation protocol to separate large protein aggregates from exosomes [ 89 ]. Indeed, even if the density of MVs remains unclear, the density of exosomes is about 1.08–1.19 g/mL [ 90 ].…”

Section: Proteomic Methodsmentioning

confidence: 99%

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

et al. 2020

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Extracellular vesicles (EVs) are lipid-bound vesicles released from cells under physiological and pathological conditions. Basing on biogenesis, dimension, content and route of secretion, they can be classified into exosomes, microvesicles (MVs) and apoptotic bodies. EVs have a key role as bioactive mediators in intercellular communication, but they are also involved in other physiological processes like immune response, blood coagulation, and tissue repair. The interest in studying EVs has increased over the years due to their involvement in several diseases, such as cardiovascular diseases (CVDs), and their potential role as biomarkers in diagnosis, therapy, and in drug delivery system development. Nowadays, the improvement of mass spectrometry (MS)-based techniques allows the characterization of the EV protein composition to deeply understand their role in several diseases. In this review, a critical overview is provided on the EV’s origin and physical properties, as well as their emerging functional role in both physiological and disease conditions, focusing attention on the role of exosomes in CVDs. The most important cardiac exosome proteomic studies will be discussed giving a qualitative and quantitative characterization of the exosomal proteins that could be used in future as new potential diagnostic markers or targets for specific therapies.

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“…Extracellular vesicles (EVs) are small lipid bilayer particles ubiquitously released by almost every cell type and present in body fluids, including urea, blood, and CSF [ 21 , 22 ]. EVs are classified into exosomes, which are secreted into the extracellular space after fusion of multivesicular bodies (MVBs) with plasma membrane [ 23 ] and microvesicles, which are created by the budding of the plasma membrane and release to the extracellular space [ 24 ], and apoptotic bodies, which are formed during the execution phase of the apoptotic process of living cells and released to the extracellular space [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Proteomic Profiling of Extracellular Vesicles Derived from Cerebrospinal Fluid of Alzheimer’s Disease Patients: A Pilot Study

Muraoka

Jedrychowski

Yanamandra

et al. 2020

Cells

Self Cite

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Pathological hallmarks of Alzheimer’s disease (AD) are deposits of amyloid beta (Aβ) and hyper-phosphorylated tau aggregates in brain plaques. Recent studies have highlighted the importance of Aβ and tau-containing extracellular vesicles (EVs) in AD. We therefore examined EVs separated from cerebrospinal fluid (CSF) of AD, mild cognitive impairment (MCI), and control (CTRL) patient samples to profile the protein composition of CSF EV. EV fractions were separated from AD (n = 13), MCI (n = 10), and CTRL (n = 10) CSF samples using MagCapture Exosome Isolation kit. The CSF-derived EV proteins were identified and quantified by label-free and tandem mass tag (TMT)-labeled mass spectrometry. Label-free proteomics analysis identified 2546 proteins that were significantly enriched for extracellular exosome ontology by Gene Ontology analysis. Canonical Pathway Analysis revealed glia-related signaling. Quantitative proteomics analysis, moreover, showed that EVs expressed 1284 unique proteins in AD, MCI and CTRL groups. Statistical analysis identified three proteins—HSPA1A, NPEPPS, and PTGFRN—involved in AD progression. In addition, the PTGFRN showed a moderate correlation with amyloid plaque (rho = 0.404, p = 0.027) and tangle scores (rho = 0.500, p = 0.005) in AD, MCI and CTRL. Based on the CSF EV proteomics, these data indicate that three proteins, HSPA1A, NPEPPS and PTGFRN, may be used to monitor the progression of MCI to AD.

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Assessment of separation methods for extracellular vesicles from human and mouse brain tissues and human cerebrospinal fluids

Cited by 49 publications

References 64 publications

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

Proteomic Profiling of Extracellular Vesicles Derived from Cerebrospinal Fluid of Alzheimer’s Disease Patients: A Pilot Study

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