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An Innovative Method for Exosome Quantification and Size Measurement

Although the biological importance of exosomes has recently gained an increasing amount of scientific and clinical attention, much is still unknown about their complex pathways, their bioavailability and their diverse functions in health and disease. Current work focuses on the presence and the beha...

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Autores principales: Mehdiani, Arash, Maier, Anatol, Pinto, Antonio, Barth, Mareike, Akhyari, Payam, Lichtenberg, Artur
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354536/
https://www.ncbi.nlm.nih.gov/pubmed/25650897
http://dx.doi.org/10.3791/50974
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author Mehdiani, Arash
Maier, Anatol
Pinto, Antonio
Barth, Mareike
Akhyari, Payam
Lichtenberg, Artur
author_facet Mehdiani, Arash
Maier, Anatol
Pinto, Antonio
Barth, Mareike
Akhyari, Payam
Lichtenberg, Artur
author_sort Mehdiani, Arash
collection PubMed
description Although the biological importance of exosomes has recently gained an increasing amount of scientific and clinical attention, much is still unknown about their complex pathways, their bioavailability and their diverse functions in health and disease. Current work focuses on the presence and the behavior of exosomes (in vitro as well as in vivo) in the context of different human disorders, especially in the fields of oncology, gynecology and cardiology. Unfortunately, neither a consensus regarding a gold standard for exosome isolation exists, nor is there an agreement on such a method for their quantitative analysis. As there are many methods for the purification of exosomes and also many possibilities for their quantitative and qualitative analysis, it is difficult to determine a combination of methods for the ideal approach. Here, we demonstrate nanoparticle tracking analysis (NTA), a semi-automated method for the characterization of exosomes after isolation from human plasma by ultracentrifugation. The presented results show that this approach for isolation, as well as the determination of the average number and size of exosomes, delivers reproducible and valid data, as confirmed by other methods, such as scanning electron microscopy (SEM).
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spelling pubmed-43545362015-03-12 An Innovative Method for Exosome Quantification and Size Measurement Mehdiani, Arash Maier, Anatol Pinto, Antonio Barth, Mareike Akhyari, Payam Lichtenberg, Artur J Vis Exp Biochemistry Although the biological importance of exosomes has recently gained an increasing amount of scientific and clinical attention, much is still unknown about their complex pathways, their bioavailability and their diverse functions in health and disease. Current work focuses on the presence and the behavior of exosomes (in vitro as well as in vivo) in the context of different human disorders, especially in the fields of oncology, gynecology and cardiology. Unfortunately, neither a consensus regarding a gold standard for exosome isolation exists, nor is there an agreement on such a method for their quantitative analysis. As there are many methods for the purification of exosomes and also many possibilities for their quantitative and qualitative analysis, it is difficult to determine a combination of methods for the ideal approach. Here, we demonstrate nanoparticle tracking analysis (NTA), a semi-automated method for the characterization of exosomes after isolation from human plasma by ultracentrifugation. The presented results show that this approach for isolation, as well as the determination of the average number and size of exosomes, delivers reproducible and valid data, as confirmed by other methods, such as scanning electron microscopy (SEM). MyJove Corporation 2015-01-17 /pmc/articles/PMC4354536/ /pubmed/25650897 http://dx.doi.org/10.3791/50974 Text en Copyright © 2015, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Biochemistry
Mehdiani, Arash
Maier, Anatol
Pinto, Antonio
Barth, Mareike
Akhyari, Payam
Lichtenberg, Artur
An Innovative Method for Exosome Quantification and Size Measurement
title An Innovative Method for Exosome Quantification and Size Measurement
title_full An Innovative Method for Exosome Quantification and Size Measurement
title_fullStr An Innovative Method for Exosome Quantification and Size Measurement
title_full_unstemmed An Innovative Method for Exosome Quantification and Size Measurement
title_short An Innovative Method for Exosome Quantification and Size Measurement
title_sort innovative method for exosome quantification and size measurement
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354536/
https://www.ncbi.nlm.nih.gov/pubmed/25650897
http://dx.doi.org/10.3791/50974
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