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Status: Bibliographieeintrag

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Verfasst von:Bordas, Marie [VerfasserIn]   i
 Genard, Géraldine [VerfasserIn]   i
 Ohl, Sibylle [VerfasserIn]   i
 Neßling, Michelle [VerfasserIn]   i
 Richter, Karsten [VerfasserIn]   i
 Roider, Tobias [VerfasserIn]   i
 Dietrich, Sascha [VerfasserIn]   i
 Maaß, Kendra K. [VerfasserIn]   i
 Seiffert, Martina [VerfasserIn]   i
Titel:Optimized protocol for isolation of small extracellular vesicles from human and murine lymphoid tissues
Verf.angabe:Marie Bordas, Géraldine Genard, Sibylle Ohl, Michelle Nessling, Karsten Richter, Tobias Roider, Sascha Dietrich, Kendra K. Maaß and Martina Seiffert
E-Jahr:2020
Jahr:4 August 2020
Fussnoten:Gesehen am 17.09.2020
Titel Quelle:Enthalten in: International journal of molecular sciences
Ort Quelle:Basel : Molecular Diversity Preservation International, 2000
Jahr Quelle:2020
Band/Heft Quelle:21(2020,15) Artikel-Nummer 5586, 15 Seiten
ISSN Quelle:1422-0067
 1661-6596
Abstract:Small extracellular vesicles (sEVs) are nanoparticles responsible for cell-to-cell communication released by healthy and cancer cells. Different roles have been described for sEVs in physiological and pathological contexts, including acceleration of tissue regeneration, modulation of tumor microenvironment, or premetastatic niche formation, and they are discussed as promising biomarkers for diagnosis and prognosis in body fluids. Although efforts have been made to standardize techniques for isolation and characterization of sEVs, current protocols often result in co-isolation of soluble protein or lipid complexes and of other extracellular vesicles. The risk of contaminated preparations is particularly high when isolating sEVs from tissues. As a consequence, the interpretation of data aiming at understanding the functional role of sEVs remains challenging and inconsistent. Here, we report an optimized protocol for isolation of sEVs from human and murine lymphoid tissues. sEVs from freshly resected human lymph nodes and murine spleens were isolated comparing two different approaches—(1) ultracentrifugation on a sucrose density cushion and (2) combined ultracentrifugation with size-exclusion chromatography. The purity of sEV preparations was analyzed using state-of-the-art techniques, including immunoblots, nanoparticle tracking analysis, and electron microscopy. Our results clearly demonstrate the superiority of size-exclusion chromatography, which resulted in a higher yield and purity of sEVs, and we show that their functionality alters significantly between the two isolation protocols.
DOI:doi:10.3390/ijms21155586
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext: https://doi.org/10.3390/ijms21155586
 Volltext: https://www.mdpi.com/1422-0067/21/15/5586
 DOI: https://doi.org/10.3390/ijms21155586
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:exosomes
 extracellular vesicles
 isolation
 lymph node
 purification
 size-exclusion chromatography
 small extracellular vesicles
 solid tissue
 spleen
 sucrose density cushion
 ultracentrifugation
K10plus-PPN:1733240586
Verknüpfungen:→ Zeitschrift

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