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Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids
Abstract
Exosomes are small membrane vesicles found in cell culture supernatants and in different biological fluids. Exosomes form in a particular population of endosomes, called multivesicular bodies (MVBs), by inward budding into the lumen of the compartment. Upon fusion of MVBs with the plasma membrane, these internal vesicles are secreted. Exosomes possess a defined set of membrane and cytosolic proteins. The physiological function of exosomes is still a matter of debate, but increasing results in various experimental systems suggest their involvement in multiple biological processes. Because both cell‐culture supernatants and biological fluids contain different types of lipid membranes, it is critical to perform high‐quality exosome purification. This unit describes different approaches for exosome purification from various sources, and discusses methods to evaluate the purity and homogeneity of the purified exosome preparations.
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“For electron microscopy (EM) analysis of EVs, samples were prepared as per the protocol described previously72.”
From GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain (2021) · cites this paper as [72]
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Metadata
- Authors
- Théry, C., Amigorena, S., Raposo, G., & Clayton, A.
- Affiliations
- Institut Curie (FR)
- Institut Curie Paris France
- Cardiff University (GB)
- Venue
- Current Protocols in Cell Biology
- Publisher
- Wiley
- Published
- 2006-03
- Volume
- 30
- Issue
- 1
- Pages
- Unit 3.22-Unit 3.22
- ISSN
- 1934-2500 (ISSN-L)
- 1934-2616
- OpenAlex
- W1877686344
- Cited by
- 5,766
- References
- 30
- Citations by year
- 2026: 206 · 2025: 356 · 2024: 485 · 2023: 502 · 2022: 558 · 2021: 641 · 2020: 620 · 2019: 521 · 2018: 452 · 2017: 390 · 2016: 301 · 2015: 259 · 2014: 177 · 2013: 106 · 2012: 85
- FWCI
- 3.54
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Current Protocols in Cell Biology (publishedVersion)
- PubMed (publishedVersion)
- ORCA Online Research @Cardiff (Cardiff University) (acceptedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Keywords
- exosomes, exosome isolation, multivesicular bodies, cell culture supernatant, biological fluids, exosome characterization, membrane vesicles, exosome function
- MeSH terms
- Animals, Culture Media, Electrophoresis, Polyacrylamide Gel, Flow Cytometry, Humans, Immunohistochemistry, Ultracentrifugation, Cell Culture Techniques, Secretory Vesicles, Microscopy, Electron, Transmission
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