Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations

Abstract Extracellular vesicles (EVs) have attracted interest due to their ability to provide diagnostic information from liquid biopsies. Cells constantly release vesicles divers in size, content and features depending on the biogenesis, origin and function. This heterogeneity adds a layer of compl...

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Autores principales:	Susann Allelein, Paula Medina-Perez, Ana Leonor Heitor Lopes, Sabrina Rau, Gerd Hause, Andreas Kölsch, Dirk Kuhlmeier
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a42e3d10d3c249a29323c9224424764c
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spelling	oai:doaj.org-article:a42e3d10d3c249a29323c9224424764c2021-12-02T15:03:05ZPotential and challenges of specifically isolating extracellular vesicles from heterogeneous populations10.1038/s41598-021-91129-y2045-2322https://doaj.org/article/a42e3d10d3c249a29323c9224424764c2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91129-yhttps://doaj.org/toc/2045-2322Abstract Extracellular vesicles (EVs) have attracted interest due to their ability to provide diagnostic information from liquid biopsies. Cells constantly release vesicles divers in size, content and features depending on the biogenesis, origin and function. This heterogeneity adds a layer of complexity when attempting to isolate and characterize EVs resulting in various protocols. Their high abundance in all bodily fluids and their stable source of origin dependent biomarkers make EVs a powerful tool in biomarker discovery and diagnostics. However, applications are limited by the quality of samples definition. Here, we compared frequently used isolation techniques: ultracentrifugation, density gradient centrifugation, ultrafiltration and size exclusion chromatography. Then, we aimed for a tissue-specific isolation of prostate-derived EVs from cell culture supernatants with immunomagnetic beads. Quality and quantity of EVs were confirmed by nanoparticle tracking analysis, western blot and electron microscopy. Additionally, a spotted antibody microarray was developed to characterize EV sub-populations. Current analysis of 16 samples on one microarray for 6 different EV surface markers in triplicate could be easily extended allowing a faster and more economical method to characterize samples.Susann AlleleinPaula Medina-PerezAna Leonor Heitor LopesSabrina RauGerd HauseAndreas KölschDirk KuhlmeierNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Susann Allelein Paula Medina-Perez Ana Leonor Heitor Lopes Sabrina Rau Gerd Hause Andreas Kölsch Dirk Kuhlmeier Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
description	Abstract Extracellular vesicles (EVs) have attracted interest due to their ability to provide diagnostic information from liquid biopsies. Cells constantly release vesicles divers in size, content and features depending on the biogenesis, origin and function. This heterogeneity adds a layer of complexity when attempting to isolate and characterize EVs resulting in various protocols. Their high abundance in all bodily fluids and their stable source of origin dependent biomarkers make EVs a powerful tool in biomarker discovery and diagnostics. However, applications are limited by the quality of samples definition. Here, we compared frequently used isolation techniques: ultracentrifugation, density gradient centrifugation, ultrafiltration and size exclusion chromatography. Then, we aimed for a tissue-specific isolation of prostate-derived EVs from cell culture supernatants with immunomagnetic beads. Quality and quantity of EVs were confirmed by nanoparticle tracking analysis, western blot and electron microscopy. Additionally, a spotted antibody microarray was developed to characterize EV sub-populations. Current analysis of 16 samples on one microarray for 6 different EV surface markers in triplicate could be easily extended allowing a faster and more economical method to characterize samples.
format	article
author	Susann Allelein Paula Medina-Perez Ana Leonor Heitor Lopes Sabrina Rau Gerd Hause Andreas Kölsch Dirk Kuhlmeier
author_facet	Susann Allelein Paula Medina-Perez Ana Leonor Heitor Lopes Sabrina Rau Gerd Hause Andreas Kölsch Dirk Kuhlmeier
author_sort	Susann Allelein
title	Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
title_short	Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
title_full	Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
title_fullStr	Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
title_full_unstemmed	Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
title_sort	potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/a42e3d10d3c249a29323c9224424764c
work_keys_str_mv	AT susannallelein potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT paulamedinaperez potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT analeonorheitorlopes potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT sabrinarau potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT gerdhause potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT andreaskolsch potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations AT dirkkuhlmeier potentialandchallengesofspecificallyisolatingextracellularvesiclesfromheterogeneouspopulations
_version_	1718389094864650240

Potential and challenges of specifically isolating extracellular vesicles from heterogeneous populations

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