Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models

Extracellular vesicles (EVs) play key roles in transporting key molecular constituents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isolation can have a profound effect on membrane integrity and yi...

Descripción completa

Detalles Bibliográficos
Autores principales: Chhoy, Peter, Brown, Caitlin W., Amante, John J., Mercurio, Arthur M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848770/
https://www.ncbi.nlm.nih.gov/pubmed/33554138
http://dx.doi.org/10.1016/j.xpro.2021.100303
_version_ 1783645198372306944
author Chhoy, Peter
Brown, Caitlin W.
Amante, John J.
Mercurio, Arthur M.
author_facet Chhoy, Peter
Brown, Caitlin W.
Amante, John J.
Mercurio, Arthur M.
author_sort Chhoy, Peter
collection PubMed
description Extracellular vesicles (EVs) play key roles in transporting key molecular constituents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isolation can have a profound effect on membrane integrity and yield. Here, we describe a step-by-step procedure to separate EVs from adherent epithelial cells using differential ultracentrifugation. Separated EVs can be further analyzed by immunoblotting, mass spectrometry, and transmission electron microscopy to derive EV yield and morphology. For complete details on the use and execution of this protocol, please refer to Brown et al. (2019).
format Online
Article
Text
id pubmed-7848770
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-78487702021-02-04 Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models Chhoy, Peter Brown, Caitlin W. Amante, John J. Mercurio, Arthur M. STAR Protoc Protocol Extracellular vesicles (EVs) play key roles in transporting key molecular constituents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isolation can have a profound effect on membrane integrity and yield. Here, we describe a step-by-step procedure to separate EVs from adherent epithelial cells using differential ultracentrifugation. Separated EVs can be further analyzed by immunoblotting, mass spectrometry, and transmission electron microscopy to derive EV yield and morphology. For complete details on the use and execution of this protocol, please refer to Brown et al. (2019). Elsevier 2021-01-29 /pmc/articles/PMC7848770/ /pubmed/33554138 http://dx.doi.org/10.1016/j.xpro.2021.100303 Text en © 2021 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Chhoy, Peter
Brown, Caitlin W.
Amante, John J.
Mercurio, Arthur M.
Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title_full Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title_fullStr Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title_full_unstemmed Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title_short Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
title_sort protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848770/
https://www.ncbi.nlm.nih.gov/pubmed/33554138
http://dx.doi.org/10.1016/j.xpro.2021.100303
work_keys_str_mv AT chhoypeter protocolfortheseparationofextracellularvesiclesbyultracentrifugationfrominvitrocellculturemodels
AT browncaitlinw protocolfortheseparationofextracellularvesiclesbyultracentrifugationfrominvitrocellculturemodels
AT amantejohnj protocolfortheseparationofextracellularvesiclesbyultracentrifugationfrominvitrocellculturemodels
AT mercurioarthurm protocolfortheseparationofextracellularvesiclesbyultracentrifugationfrominvitrocellculturemodels

Ejemplares similares