A novel strategy for facile serum exosome isolation based on specific interactions between phospholipid bilayers and TiO<sub>2</sub>

Gao, Fangyuan; Jiao, Fenglong; Xia, Chaoshuang; Zhao, Yang; Ying, Wantao; Xie, Yunying; Guan, Xiaoya; Tao, Ming; Zhang, Yangjun; Qin, Weijie; Qian, Xiaohong

doi:10.1039/c8sc04197k

Cited by 165 publications

(116 citation statements)

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“…However, an affinity for the lipid membrane structures of EVs is often applied to facilitate plasma EV extraction. The enrichment of phosphopeptides on the lipid bilayer of EVs by titanium dioxide (TiO 2 ) affinity chromatography allowed a rapid and efficient isolation of high-quality exosomes from human serum of pancreatic cancer patients and healthy subjects [ 112 ]. Besides, EVs can be purified based on their membrane phospholipid composition, in particular phosphatidylserines and GM1 gangliosides.…”

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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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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“…EVs were also isolated from the WKY-AF and SHR-AF medium with ultracentrifugation method [32]t o confirm the effects of EVs that isolated with the PureExo® Exosome Isolation Kit. Briefly, the collect medium was centrifuged at 300 × g for 5 min, 3000 × g for 30 min and then 10,000 × g for 60 min to remove dead cells, cell debris, apoptotic bodies and large vesicles (-500-1000 nm).…”

Section: Isolation Of Evs From Shr-af and Wky-af Culturesmentioning

confidence: 99%

MiR155‐5p in adventitial fibroblasts‐derived extracellular vesicles inhibits vascular smooth muscle cell proliferation via suppressing angiotensin‐converting enzyme expression

Ren

Tong

Qiu

et al. 2019

J of Extracellular Vesicle

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2020) MiR155-5p in adventitial fibroblastsderived extracellular vesicles inhibits vascular smooth muscle cell proliferation via suppressing ABSTRACT Proliferation of vascular smooth muscle cells (VSMCs) plays crucial roles in vascular remodelling and stiffening in hypertension. Vascular adventitial fibroblasts are a key regulator of vascular wall function and structure. This study is designed to investigate the roles of adventitial fibroblasts-derived extracellular vesicles (EVs) in VSMC proliferation and vascular remodelling in normotensive Wistar-Kyoto rat (WKY) and spontaneously hypertensive rat (SHR), an animal model of human essential hypertension. EVs were isolated from aortic adventitial fibroblasts of WKY (WKY-EVs) and SHR (SHR-EVs). Compared with WKY-EVs, miR155-5p content was reduced, while angiotensin-converting enzyme (ACE) content was increased in SHR-EVs. WKY-EVs inhibited VSMC proliferation of SHR, which was prevented by miR155-5p inhibitor. SHR-EVs promoted VSMC proliferation of both strains, which was enhanced by miR155-5p inhibitor, but abolished by captopril or losartan. Dual luciferase reporter assay showed that ACE was a target gene of miR155-5p. MiR155-5p mimic or overexpression inhibited VSMC proliferation and ACE upregulation of SHR. WKY-EVs reduced ACE mRNA and protein expressions while SHR-EVs only increased ACE protein level in VSMCs of both strains. However, the SHR-EVs-derived from the ACE knockdown-treated adventitial fibroblasts lost the roles in promoting VSMC proliferation and ACE upregulation. Systemic miR155-5p overexpression reduced vascular ACE, angiotensin II and proliferating cell nuclear antigen levels, and attenuated hypertension and vascular remodelling in SHR. Repetitive intravenous injection of SHR-EVs increased blood pressure and vascular ACE contents, and promoted vascular remodelling in both strains, while WKY-EVs reduced vascular ACE contents and attenuated hypertension and vascular remodelling in SHR. We concluded that WKY-EVs-mediated miR155-5p transfer attenuates VSMC proliferation and vascular remodelling in SHR via suppressing ACE expression, while SHR-EVs-mediated ACE transfer promotes VSMC proliferation and vascular remodelling. ARTICLE HISTORY

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“…Exosomes are 30–150 nm lipid bilayer‐enclosed vesicles released to the extracellular space after the fusion of intracellular polyvesicles and cell membranes . Almost all cells can produce exosomes, and they can be found in all biological fluids including serum, plasma, urine, saliva, amniotic fluid, semen, and cell cultured medium .…”

Section: Protein Ms/ms Quantity Of the Known Exosomal Proteins And Comentioning

confidence: 99%

Proteomics comparison of exosomes from serum and plasma between ultracentrifugation and polymer‐based precipitation kit methods

et al. 2019

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Exosomes are vesicles with sizes ranging from 30 to 150 nm. The analysis and detection of blood exosomes offers an effective route for cancer diagnosis, prognosis assessment, and therapeutic evaluation of diseases. Due to the difference in separation procedure, collection method and the usage of anticoagulants, serum and plasma samples show diversity test results. In order to evaluate the isolation effect of exosomes in serum and plasma samples, two commonly used exosomal isolation methods, ultracentrifugation and polymer‐based precipitation kit, were used, respectively. And the isolation effects were evaluated by comparing the composition and abundant of proteins from isolated exosomes based on MS‐based proteomics analysis. The results showed that the plasma exosomes extracted by ultracentrifugation identified more exosome biomarkers, and the concentrations of these biomarkers were higher than others. And plasma exosomes could be a better sample for blood‐based proteomics research of exosomes. It would be more useful for future targeted biomarker discovery.

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A novel strategy for facile serum exosome isolation based on specific interactions between phospholipid bilayers and TiO₂

Abstract: Exosomes can be efficiently isolated in a short period of time by the specific interaction of titanium dioxide with the phosphate groups on the surface of phospholipid bilayer.

Cited by 165 publications

References 50 publications

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

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

MiR155‐5p in adventitial fibroblasts‐derived extracellular vesicles inhibits vascular smooth muscle cell proliferation via suppressing angiotensin‐converting enzyme expression

Proteomics comparison of exosomes from serum and plasma between ultracentrifugation and polymer‐based precipitation kit methods

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A novel strategy for facile serum exosome isolation based on specific interactions between phospholipid bilayers and TiO2

Abstract: Exosomes can be efficiently isolated in a short period of time by the specific interaction of titanium dioxide with the phosphate groups on the surface of phospholipid bilayer.

Cited by 165 publications

References 50 publications

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

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

MiR155‐5p in adventitial fibroblasts‐derived extracellular vesicles inhibits vascular smooth muscle cell proliferation via suppressing angiotensin‐converting enzyme expression

Proteomics comparison of exosomes from serum and plasma between ultracentrifugation and polymer‐based precipitation kit methods

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A novel strategy for facile serum exosome isolation based on specific interactions between phospholipid bilayers and TiO₂