Magnetic Bead-Based Isolation of Exosomes

Oksvold, Morten P.; Neurauter, Axl; Pedersen, Ketil W.

doi:10.1007/978-1-4939-1538-5_27

Cited by 160 publications

(119 citation statements)

References 15 publications

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“…To map surface epitopes, we used a previously published method (35) whereby EVs are first trapped with a large bead linked to an antibody to CD63, and then additional epitopes on the trapped EVs are assayed with standard protocols for flow cytometry. As expected, the EVs captured with the protocol were positive for CD63 (Fig.…”

Section: Selection Of Optimal Mscs and Culture Conditions For Productmentioning

confidence: 99%

Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Kim¹,

Nishida²,

An³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

399

325

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Extracellular vesicles (EVs) secreted by cells present an attractive strategy for developing new therapies, but progress in the field is limited by several issues: The quality of the EVs varies with the type and physiological status of the producer cells; protocols used to isolate the EVs are difficult to scale up; and assays for efficacy are difficult to develop. In the present report, we have addressed these issues by using human mesenchymal stem/stromal cells (MSCs) that produce EVs when incubated in a protein-free medium, preselecting the preparations of MSCs with a biomarker for their potency in modulating inflammation, incubating the cells in a chemically defined protein-free medium that provided a stable environment, isolating the EVs with a scalable chromatographic procedure, and developing an in vivo assay for efficacy of the cells in suppressing neuroinflammation after traumatic brain injury (TBI) in mice. In addition, we demonstrate that i.v. infusion of the isolated EVs shortly after induction of TBI rescued pattern separation and spatial learning impairments 1 mo later.MSCs | neuroinflammation | exosomes | efficacy assay

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Section: Selection Of Optimal Mscs and Culture Conditions For Productmentioning

confidence: 99%

Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Kim¹,

Nishida²,

An³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

399

325

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“…Several methods of isolation have been described ranging from ultracentrifugation, density gradient centrifugation, immunoaffinity capture using magnetic beads and commercially available precipitation methods. Comparative studies using these techniques demonstrated that the purity and the quality of preparations is dependent on the source of exosomes …”

Section: Extracellular Vesicles— Isolationmentioning

confidence: 99%

Extracellular vesicles as carriers of microRNA, proteins and lipids in tumor microenvironment

et al. 2015

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In recent years, the knowledge about the control of tumor microenvironment has increased and emerged as an important player in tumorigenesis. The role of normal stromal cells in the tumor initiation and progression has brought our vision in to the forefront of cell-to-cell communication. In this review, we focus on the mechanism of communication between stromal and tumor cells, which is based on the exchange of extracellular vesicles (EVs). We describe several, evergrowing, pieces of evidence that EVs transfer messages through their miRNA, lipid, protein and nucleic acid contents. A better understanding of this sophisticated method of communication between normal cancer cells may lead to developing novel approaches for personalized diagnostics and therapeutics.

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“…In this regard, choosing a proper exosome membrane marker is one of the most important steps in these immunoassays. It was shown that members of the tetraspanin family, such as CD81, CD9, and CD63, that are present on the membranes of exosomes can be used for efficient immunocapture . Unlike other conventional techniques for exosome isolation, antibody‐coated magnetic beads can be used for direct exosome isolation from bodily fluids, which reduces the time‐consuming centrifugation steps.…”

Section: Isolation Of Exosomesmentioning

confidence: 99%

Biological Functions and Current Advances in Isolation and Detection Strategies for Exosome Nanovesicles

Boriachek

Islam

Möller

et al. 2017

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Exosomes are nanoscale (≈30-150 nm) extracellular vesicles of endocytic origin that are shed by most types of cells and circulate in bodily fluids. Exosomes carry a specific composition of proteins, lipids, RNA, and DNA and can work as cargo to transfer this information to recipient cells. Recent studies on exosomes have shown that they play an important role in various biological processes, such as intercellular signaling, coagulation, inflammation, and cellular homeostasis. These functional roles are attributed to their ability to transfer RNA, proteins, enzymes, and lipids, thereby affecting the physiological and pathological conditions in various diseases, including cancer and neurodegenerative, infectious, and autoimmune diseases (e.g., cancer initiation, progression, and metastasis). Due to these unique characteristics, exosomes are considered promising biomarkers for the diagnosis and prognosis of various diseases via noninvasive or minimally invasive procedures. Over the last decade, a plethora of methodologies have been developed for analyzing disease-specific exosomes using optical and nonoptical tools. Here, the major biological functions, significance, and potential role of exosomes as biomarkers and therapeutics are discussed. Furthermore, an overview of the most commonly used techniques for exosome analysis, highlighting the major technical challenges and limitations of existing techniques, is presented.

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Magnetic Bead-Based Isolation of Exosomes

Cited by 160 publications

References 15 publications

Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Extracellular vesicles as carriers of microRNA, proteins and lipids in tumor microenvironment

Biological Functions and Current Advances in Isolation and Detection Strategies for Exosome Nanovesicles

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Magnetic Bead-Based Isolation of Exosomes

Cited by 160 publications

References 15 publications

Chromatographically isolated CD63 + CD81 + extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Chromatographically isolated CD63 + CD81 + extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Extracellular vesicles as carriers of microRNA, proteins and lipids in tumor microenvironment

Biological Functions and Current Advances in Isolation and Detection Strategies for Exosome Nanovesicles

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Product

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Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI

Chromatographically isolated CD63 ⁺ CD81 ⁺ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI