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Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences

[Image: see text] Microglial cells are a component of the innate immune system in the brain that support cell-to-cell communication via secreted molecules and extracellular vesicles (EVs). EVs can be divided into two major populations: large (LEVs) and small (SEVs) EVs, carrying different mediators,...

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Autores principales: Ceccarelli, Lorenzo, Marchetti, Laura, Rizzo, Milena, Moscardini, Aldo, Cappello, Valentina, Da Pozzo, Eleonora, Romano, Miriam, Giacomelli, Chiara, Bergese, Paolo, Martini, Claudia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9280972/
https://www.ncbi.nlm.nih.gov/pubmed/35847267
http://dx.doi.org/10.1021/acsomega.2c00816
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author Ceccarelli, Lorenzo
Marchetti, Laura
Rizzo, Milena
Moscardini, Aldo
Cappello, Valentina
Da Pozzo, Eleonora
Romano, Miriam
Giacomelli, Chiara
Bergese, Paolo
Martini, Claudia
author_facet Ceccarelli, Lorenzo
Marchetti, Laura
Rizzo, Milena
Moscardini, Aldo
Cappello, Valentina
Da Pozzo, Eleonora
Romano, Miriam
Giacomelli, Chiara
Bergese, Paolo
Martini, Claudia
author_sort Ceccarelli, Lorenzo
collection PubMed
description [Image: see text] Microglial cells are a component of the innate immune system in the brain that support cell-to-cell communication via secreted molecules and extracellular vesicles (EVs). EVs can be divided into two major populations: large (LEVs) and small (SEVs) EVs, carrying different mediators, such as proteins, lipids, and miRNAs. The microglia EVs cargo crucially reflects the status of parental cells and can lead to both beneficial and detrimental effects in many physiopathological states. Herein, a workflow for the extraction and characterization of SEVs and LEVs from human C20 and HMC3 microglia cell lines derived, respectively, from adult and embryonic microglia is reported. EVs were gathered from the culture media of the two cell lines by sequential ultracentrifugation steps and their biochemical and biophysical properties were analyzed by Western blot, transmission electron microscopy, and dynamic light scattering. Although the C20- and HMC3-derived EVs shared several common features, C20-derived EVs were slightly lower in number and more polydispersed. Interestingly, C20- but not HMC3-SEVs were able to interfere with the proliferation of U87 glioblastoma cells. This correlated with the different relative levels of eight miRNAs involved in neuroinflammation and tumor progression in the C20- and HMC3-derived EVs, which in turn reflected a different basal activation state of the two cell types. Our data fill a gap in the community of microglia EVs, in which the preparations from human cells have been poorly characterized so far. Furthermore, these results shed light on both the differences and similarities of EVs extracted from different human microglia cell models, underlining the need to better characterize the features and biological effects of EVs for therein useful and correct application.
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spelling pubmed-92809722022-07-15 Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences Ceccarelli, Lorenzo Marchetti, Laura Rizzo, Milena Moscardini, Aldo Cappello, Valentina Da Pozzo, Eleonora Romano, Miriam Giacomelli, Chiara Bergese, Paolo Martini, Claudia ACS Omega [Image: see text] Microglial cells are a component of the innate immune system in the brain that support cell-to-cell communication via secreted molecules and extracellular vesicles (EVs). EVs can be divided into two major populations: large (LEVs) and small (SEVs) EVs, carrying different mediators, such as proteins, lipids, and miRNAs. The microglia EVs cargo crucially reflects the status of parental cells and can lead to both beneficial and detrimental effects in many physiopathological states. Herein, a workflow for the extraction and characterization of SEVs and LEVs from human C20 and HMC3 microglia cell lines derived, respectively, from adult and embryonic microglia is reported. EVs were gathered from the culture media of the two cell lines by sequential ultracentrifugation steps and their biochemical and biophysical properties were analyzed by Western blot, transmission electron microscopy, and dynamic light scattering. Although the C20- and HMC3-derived EVs shared several common features, C20-derived EVs were slightly lower in number and more polydispersed. Interestingly, C20- but not HMC3-SEVs were able to interfere with the proliferation of U87 glioblastoma cells. This correlated with the different relative levels of eight miRNAs involved in neuroinflammation and tumor progression in the C20- and HMC3-derived EVs, which in turn reflected a different basal activation state of the two cell types. Our data fill a gap in the community of microglia EVs, in which the preparations from human cells have been poorly characterized so far. Furthermore, these results shed light on both the differences and similarities of EVs extracted from different human microglia cell models, underlining the need to better characterize the features and biological effects of EVs for therein useful and correct application. American Chemical Society 2022-06-28 /pmc/articles/PMC9280972/ /pubmed/35847267 http://dx.doi.org/10.1021/acsomega.2c00816 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Ceccarelli, Lorenzo
Marchetti, Laura
Rizzo, Milena
Moscardini, Aldo
Cappello, Valentina
Da Pozzo, Eleonora
Romano, Miriam
Giacomelli, Chiara
Bergese, Paolo
Martini, Claudia
Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title_full Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title_fullStr Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title_full_unstemmed Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title_short Human Microglia Extracellular Vesicles Derived from Different Microglia Cell Lines: Similarities and Differences
title_sort human microglia extracellular vesicles derived from different microglia cell lines: similarities and differences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9280972/
https://www.ncbi.nlm.nih.gov/pubmed/35847267
http://dx.doi.org/10.1021/acsomega.2c00816
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