Isolation of Platelet-Derived Extracellular Vesicles

Aatonen, Maria; Valkonen, Sami; Böing, Anita N.; Yuana, Yuana; Nieuwland, Rienk; Siljander, Pia

doi:10.1007/978-1-4939-6728-5_12

Cited by 25 publications

(25 citation statements)

References 15 publications

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“…In fact, the formation of these clumps or aggregates probably explains why previously, on relative insensitive flow cytometers, mainly "EVs" of platelet origin were measured [2,15]. Given the fact that recent evidence suggests that the bulk of small EVs cannot be pelleted from plasma or serum by (ultra) centrifugation supports the idea that residual platelets rather than platelet-derived EVs explained these earlier observations [8].…”

Section: Discussionmentioning

confidence: 86%

Extracellular vesicles and coagulation in blood from healthy humans revisited

Berckmans

Lacroix

Hau

et al. 2019

J of Extracellular Vesicle

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Background: In 2001, we studied the presence and coagulant properties of "microparticles" in the blood of healthy humans. Since then, multiple improvements in detection, isolation and functional characterization of the now called "extracellular vesicles" (EVs) have been made, and shortcomings were identified. Aim: To revisit the presence and function of EVs in blood from healthy humans. Methods: Blood was collected from 20 healthy donors. EV-containing plasma was prepared according to new guidelines, and plasma was diluted to prevent swarm detection. Single EVs were measured by flow cytometry with known sensitivity of fluorescence and light scatter. The haemostatic properties of EVs were measured by thrombin-, fibrin-, and plasmin generation. Plasma concentrations of thrombin-antithrombin complexes and prothrombin fragment 1 + 2 were measured to assess the coagulation status in vivo. Results: Compared to 2001, the total concentrations of detected EVs increased from 190-to 264-fold. In contrast to 2001, however, EVs are non-coagulant which we show can be attributed to improvements in blood collection and plasma preparation. No relation is present between the plasma concentrations of EVs and either TAT or F1 + 2. Finally, we show that EVs support plasmin generation. Discussion: Improvements in blood collection, plasma preparation and detection of EVs reveal that results from earlier studies have to be interpreted with care. Compared to 2001, higher concentrations of EVs are detected in blood of healthy humans which promote fibrinolysis rather than coagulation.

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Section: Discussionmentioning

confidence: 86%

Extracellular vesicles and coagulation in blood from healthy humans revisited

Berckmans

Lacroix

Hau

et al. 2019

J of Extracellular Vesicle

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“…Negative selection with magnetic beads was performed for preparation of Exos‐depleted CM (EDCM) considering its relatively negligible effect on the remaining components of CM after magnetic bead‐based Exos depletion. [ 30 ]…”

Section: Methodsmentioning

confidence: 99%

EWSAT1 Acts in Concert with Exosomes in Osteosarcoma Progression and Tumor‐Induced Angiogenesis: The “Double Stacking Effect”

et al. 2020

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The prognosis for osteosarcoma (OS) continues to be unsatisfactory due to tumor recurrence as a result of metastasis and drug resistance. Several studies have shown that Ewing sarcoma associated transcript 1 (EWSAT1) plays an important role in the progression of OS. Exosomes (Exos) act as important carriers in intercellular communication and play an important role in the tumor microenvironment, especially in tumor‐induced angiogenesis. Nonetheless, the specific mechanism via which EWSAT1 and Exos regulate OS progression is unknown, and whether they can be effective therapeutic targets also requires verification. Hence, in this study, it is aimed to investigate the mechanisms of action of EWSAT1 and Exos. EWSAT1 significantly promotes proliferation, migration, colony formation, and survival of OS. EWSAT1 regulates OS‐induced angiogenesis via two mechanisms, called the “double stacking effect,” which is a combination of the increase in sensitivity/reactivity of vascular endothelial cells triggered by Exos‐carrying EWSAT1, and the EWSAT1‐induced increase in angiogenic factor secretion. In vivo experiments further validates the “double stacking effect” and shows that EWSAT1‐KD effectively inhibits tumor growth in OS. The above observations indicate that EWSAT1 can be used as not only a potential diagnostic and prognostic marker, but also as a precise therapeutic target for OS.

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“…It has been found that upon activation, blood platelets are able to release two types of EVs, i.e., microparticles and exosomes [34,35] . Platelet-derived microparticles (also referred to microvesicles) are characterized by an average of the size of 70-100 nm and the PS presentation at their surface, which mediates their capabilities in coagulation and thrombosis [36] . On contrary, platelet-derived exosomes are characterized by their small size (< 80 nm) and the presence of CD63 antigen, which is known as the tetraspanin that belongs to the transmembrane 4 superfamily, while larger vesicles (> 0.5 µm) containing chromatin materials are referred to apoptotic bodies [35,36] .…”

Section: Platelet-derived Vesiclesmentioning

confidence: 99%

Platelet-derived vesicles: diagnostic and predictive value in cardiovascular diseases

Berezin¹,

Berezin²

2019

JUMD

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There is emerging evidence that endogenous microvesicles released by platelets may play a pivotal role in pathogenesis of numerous diseases including stable and unstable ischemic heart disease, atherosclerosis, hypertension, idiopathic pulmonary hypertension, atrial fibrillation/flutter, acute and chronic heart failure, as well as systemic vasculitis, systemic lupus erythematosus, antiphospholipid syndrome, transplant rejection, diabetes-induced angiopathy. Microvesicles that leased from the platelets exist as cargo for a large of number biological-active molecules including regulatory peptides, hormones, growth factors, active molecules coordinating cell-to-cell cooperation. Moreover, platelet-derived microvesicles are concerned about a conspicuous component in regulating angiogenesis, cardiac protection, and vascular repair. Recent animal and clinical studies have shown that the number of circulating platelet-derived microvesicles may sufficiently increase in accelerating atherosclerosis, acute atherothrombotic events, i.e., recurrent ischemia, myocardial infarction, thrombosis, hypertension, and stroke, microvascular inflammation. It is suggested that exaggerating levels of circulating platelet-derived microvesicles is strongly associated with endothelial dysfunction. The review is discussed the role of platelet-derived vesicles in the pathogenesis of the cardiovascular disease, as well as diagnostic and predictive capabilities of platelet-derived vesicles as biomarkers of a natural evolution of the diseases.

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Isolation of Platelet-Derived Extracellular Vesicles

Cited by 25 publications

References 15 publications

Extracellular vesicles and coagulation in blood from healthy humans revisited

Extracellular vesicles and coagulation in blood from healthy humans revisited

EWSAT1 Acts in Concert with Exosomes in Osteosarcoma Progression and Tumor‐Induced Angiogenesis: The “Double Stacking Effect”

Platelet-derived vesicles: diagnostic and predictive value in cardiovascular diseases

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