Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture
Summary: Extracellular vesicles (EVs) play important roles in extracellular trafficking and signaling. Here, we separate EVs by differential centrifugation. EVs separated by this approach are called large EVs (l-EVs) and small EVs (s-EVs), reflecting particle size, which sediment based on different...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721003324 |
| _version_ | 1818360988661972992 |
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| author | Hooi Ting Hu Tamako Nishimura Shiro Suetsugu |
| author_facet | Hooi Ting Hu Tamako Nishimura Shiro Suetsugu |
| author_sort | Hooi Ting Hu |
| collection | DOAJ |
| description | Summary: Extracellular vesicles (EVs) play important roles in extracellular trafficking and signaling. Here, we separate EVs by differential centrifugation. EVs separated by this approach are called large EVs (l-EVs) and small EVs (s-EVs), reflecting particle size, which sediment based on different ultracentrifugation forces. The resulting EVs can be quantified and analyzed using nanoparticle tracking analysis, immunoblotting, and functional assays. This protocol was applied to a suspension cell line with high transfection efficiency adapted to a high-density, serum-free culture.For complete details on the use and execution of this protocol, please refer to Nishimura et al. (2021). |
| first_indexed | 2024-12-13T21:09:32Z |
| format | Article |
| id | doaj.art-0ebebf1f04cc41de806f0915956c49e3 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-13T21:09:32Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-0ebebf1f04cc41de806f0915956c49e32022-12-21T23:31:23ZengElsevierSTAR Protocols2666-16672021-09-0123100625Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell cultureHooi Ting Hu0Tamako Nishimura1Shiro Suetsugu2Division of Biological Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, Ikoma 630-0192, Japan; Corresponding authorDivision of Biological Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, Ikoma 630-0192, JapanDivision of Biological Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, Ikoma 630-0192, Japan; Data Science Center, Nara Institute of Science and Technology, Ikoma 630-0192, Japan; Center for Digital Green-innovation, Nara Institute of Science and Technology, Ikoma 630-0192, Japan; Corresponding authorSummary: Extracellular vesicles (EVs) play important roles in extracellular trafficking and signaling. Here, we separate EVs by differential centrifugation. EVs separated by this approach are called large EVs (l-EVs) and small EVs (s-EVs), reflecting particle size, which sediment based on different ultracentrifugation forces. The resulting EVs can be quantified and analyzed using nanoparticle tracking analysis, immunoblotting, and functional assays. This protocol was applied to a suspension cell line with high transfection efficiency adapted to a high-density, serum-free culture.For complete details on the use and execution of this protocol, please refer to Nishimura et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166721003324Cell cultureCell MembraneCell separation/fractionationMolecular BiologyAntibodySignal Transduction |
| spellingShingle | Hooi Ting Hu Tamako Nishimura Shiro Suetsugu Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture STAR Protocols Cell culture Cell Membrane Cell separation/fractionation Molecular Biology Antibody Signal Transduction |
| title | Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture |
| title_full | Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture |
| title_fullStr | Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture |
| title_full_unstemmed | Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture |
| title_short | Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture |
| title_sort | ultracentrifugal separation characterization and functional study of extracellular vesicles derived from serum free cell culture |
| topic | Cell culture Cell Membrane Cell separation/fractionation Molecular Biology Antibody Signal Transduction |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721003324 |
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