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Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models

The role of extracellular vesicles (EVs) has been completely re-evaluated in the recent decades, and EVs are currently considered to be among the main players in intercellular communication. Beyond their functional aspects, there is strong interest in the development of faster and less expensive iso...

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Autores principales: Serratì, Simona, Palazzo, Antonio, Lapenna, Annamaria, Mateos, Helena, Mallardi, Antonia, Marsano, René Massimiliano, Quarta, Alessandra, Del Rosso, Mario, Azzariti, Amalia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699204/
https://www.ncbi.nlm.nih.gov/pubmed/34944501
http://dx.doi.org/10.3390/biom11121857
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author Serratì, Simona
Palazzo, Antonio
Lapenna, Annamaria
Mateos, Helena
Mallardi, Antonia
Marsano, René Massimiliano
Quarta, Alessandra
Del Rosso, Mario
Azzariti, Amalia
author_facet Serratì, Simona
Palazzo, Antonio
Lapenna, Annamaria
Mateos, Helena
Mallardi, Antonia
Marsano, René Massimiliano
Quarta, Alessandra
Del Rosso, Mario
Azzariti, Amalia
author_sort Serratì, Simona
collection PubMed
description The role of extracellular vesicles (EVs) has been completely re-evaluated in the recent decades, and EVs are currently considered to be among the main players in intercellular communication. Beyond their functional aspects, there is strong interest in the development of faster and less expensive isolation protocols that are as reliable for post-isolation characterisations as already-established methods. Therefore, the identification of easy and accessible EV isolation techniques with a low price/performance ratio is of paramount importance. We isolated EVs from a wide spectrum of samples of biological and clinical interest by choosing two isolation techniques, based on their wide use and affordability: ultracentrifugation and salting-out. We collected EVs from human cancer and healthy cell culture media, yeast, bacteria and Drosophila culture media and human fluids (plasma, urine and saliva). The size distribution and concentration of EVs were measured by nanoparticle tracking analysis and dynamic light scattering, and protein depletion was measured by a colorimetric nanoplasmonic assay. Finally, the EVs were characterised by flow cytometry. Our results showed that the salting-out method had a good efficiency in EV separation and was more efficient in protein depletion than ultracentrifugation. Thus, salting-out may represent a good alternative to ultracentrifugation.
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spelling pubmed-86992042021-12-24 Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models Serratì, Simona Palazzo, Antonio Lapenna, Annamaria Mateos, Helena Mallardi, Antonia Marsano, René Massimiliano Quarta, Alessandra Del Rosso, Mario Azzariti, Amalia Biomolecules Article The role of extracellular vesicles (EVs) has been completely re-evaluated in the recent decades, and EVs are currently considered to be among the main players in intercellular communication. Beyond their functional aspects, there is strong interest in the development of faster and less expensive isolation protocols that are as reliable for post-isolation characterisations as already-established methods. Therefore, the identification of easy and accessible EV isolation techniques with a low price/performance ratio is of paramount importance. We isolated EVs from a wide spectrum of samples of biological and clinical interest by choosing two isolation techniques, based on their wide use and affordability: ultracentrifugation and salting-out. We collected EVs from human cancer and healthy cell culture media, yeast, bacteria and Drosophila culture media and human fluids (plasma, urine and saliva). The size distribution and concentration of EVs were measured by nanoparticle tracking analysis and dynamic light scattering, and protein depletion was measured by a colorimetric nanoplasmonic assay. Finally, the EVs were characterised by flow cytometry. Our results showed that the salting-out method had a good efficiency in EV separation and was more efficient in protein depletion than ultracentrifugation. Thus, salting-out may represent a good alternative to ultracentrifugation. MDPI 2021-12-10 /pmc/articles/PMC8699204/ /pubmed/34944501 http://dx.doi.org/10.3390/biom11121857 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Serratì, Simona
Palazzo, Antonio
Lapenna, Annamaria
Mateos, Helena
Mallardi, Antonia
Marsano, René Massimiliano
Quarta, Alessandra
Del Rosso, Mario
Azzariti, Amalia
Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title_full Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title_fullStr Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title_full_unstemmed Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title_short Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models
title_sort salting-out approach is worthy of comparison with ultracentrifugation for extracellular vesicle isolation from tumor and healthy models
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699204/
https://www.ncbi.nlm.nih.gov/pubmed/34944501
http://dx.doi.org/10.3390/biom11121857
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