Isolation and characterization of extracellular vesicle subpopulations from tissues. Crescitelli, R., Lässer, C., & Lötvall, J. Nature Protocols, 16(3):1548–1580, March, 2021. Number: 3 Publisher: Nature Publishing Group![Isolation and characterization of extracellular vesicle subpopulations from tissues [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex Extracellular vesicles (EVs) are lipid bilayered membrane structures released by all cells. Most EV studies have been performed by using cell lines or body fluids, but the number of studies on tissue-derived EVs is still limited. Here, we present a protocol to isolate up to six different EV subpopulations directly from tissues. The approach includes enzymatic treatment of dissociated tissues followed by differential ultracentrifugation and density separation. The isolated EV subpopulations are characterized by electron microscopy and RNA profiling. In addition, their protein cargo can be determined with mass spectrometry, western blot and ExoView. Tissue-EV isolation can be performed in 22 h, but a simplified version can be completed in 8 h. Most experiments with the protocol have used human melanoma metastases, but the protocol can be applied to other cancer and non-cancer tissues. The procedure can be adopted by researchers experienced with cell culture and EV isolation.
@article{crescitelli_isolation_2021,
title = {Isolation and characterization of extracellular vesicle subpopulations from tissues},
volume = {16},
copyright = {2021 The Author(s), under exclusive licence to Springer Nature Limited},
issn = {1750-2799},
url = {https://www.nature.com/articles/s41596-020-00466-1},
doi = {10.1038/s41596-020-00466-1},
abstract = {Extracellular vesicles (EVs) are lipid bilayered membrane structures released by all cells. Most EV studies have been performed by using cell lines or body fluids, but the number of studies on tissue-derived EVs is still limited. Here, we present a protocol to isolate up to six different EV subpopulations directly from tissues. The approach includes enzymatic treatment of dissociated tissues followed by differential ultracentrifugation and density separation. The isolated EV subpopulations are characterized by electron microscopy and RNA profiling. In addition, their protein cargo can be determined with mass spectrometry, western blot and ExoView. Tissue-EV isolation can be performed in 22 h, but a simplified version can be completed in 8 h. Most experiments with the protocol have used human melanoma metastases, but the protocol can be applied to other cancer and non-cancer tissues. The procedure can be adopted by researchers experienced with cell culture and EV isolation.},
language = {en},
number = {3},
urldate = {2023-04-07},
journal = {Nature Protocols},
author = {Crescitelli, Rossella and Lässer, Cecilia and Lötvall, Jan},
month = mar,
year = {2021},
note = {Number: 3
Publisher: Nature Publishing Group},
keywords = {Cancer microenvironment, Cell signalling, Isolation, Membrane trafficking, separation and purification},
pages = {1548--1580},
}
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