| |  Online-Ressource |
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| Verfasst von: | Bordas, Marie [VerfasserIn]  |
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| | Genard, Géraldine [VerfasserIn] |
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| | Ohl, Sibylle [VerfasserIn] |
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| | Neßling, Michelle [VerfasserIn] |
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| | Richter, Karsten [VerfasserIn] |
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| | Roider, Tobias [VerfasserIn] |
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| | Dietrich, Sascha [VerfasserIn] |
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| | Maaß, Kendra K. [VerfasserIn] |
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| | Seiffert, Martina [VerfasserIn] |
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| Titel: | Optimized protocol for isolation of small extracellular vesicles from human and murine lymphoid tissues |
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| Verf.angabe: | Marie Bordas, Géraldine Genard, Sibylle Ohl, Michelle Nessling, Karsten Richter, Tobias Roider, Sascha Dietrich, Kendra K. Maaß and Martina Seiffert |
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| E-Jahr: | 2020 |
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| Jahr: | 4 August 2020 |
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| Fussnoten: | Gesehen am 17.09.2020 |
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| Titel Quelle: | Enthalten in: International journal of molecular sciences |
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| Ort Quelle: | Basel : Molecular Diversity Preservation International, 2000 |
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| Jahr Quelle: | 2020 |
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| Band/Heft Quelle: | 21(2020,15) Artikel-Nummer 5586, 15 Seiten |
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| ISSN Quelle: | 1422-0067 |
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| | 1661-6596 |
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| 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. |
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| DOI: | doi:10.3390/ijms21155586 |
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| 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 |
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| | Volltext: https://www.mdpi.com/1422-0067/21/15/5586 |
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| | DOI: https://doi.org/10.3390/ijms21155586 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | exosomes |
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| | extracellular vesicles |
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| | isolation |
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| | lymph node |
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| | purification |
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| | size-exclusion chromatography |
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| | small extracellular vesicles |
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| | solid tissue |
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| | spleen |
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| | sucrose density cushion |
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| | ultracentrifugation |
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| K10plus-PPN: | 1733240586 |
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| Verknüpfungen: | → Zeitschrift |
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Optimized protocol for isolation of small extracellular vesicles from human and murine lymphoid tissues / Bordas, Marie [VerfasserIn]; 4 August 2020 (Online-Ressource)
