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Isolation and characterization of exosomes from cultures of tissue explants and cell lines
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Clinical Immunology.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Clinical Immunology.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Clinical Immunology.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Clinical Immunology.
2016 (English)In: Current Protocols in Immunology, ISSN 1934-3671, Vol. 2016, p. 14.42.1-14.42.21Article in journal (Refereed) Published
Abstract [en]

Exosomes are specialized, nanometer-sized extracellular vesicles of endosomal origin actively secreted into the extracellular space by a variety of cells under normal and pathological conditions. Exosomes have recently emerged as important intercellular communicators and modulators of diverse mechanisms and cellular responses. Characterization of their composition and functionwill open possibilities for new diagnostic methods and promising therapeutic approaches based on nanobiology. This unit provides a standard isolation procedure for purification of exosomes based on density gradient ultracentrifugation with sucrose. The process of isolating exosomes relies on obtaining proper source fluids/supernatants as well as qualitative and quantitative assessment of the isolated vesicles. The methodological procedures here can be divided in three parts: (1) pre-isolation procedures aiming to obtain fluids containing exosomes, with a focus on protocols for organ explants and cell cultures; (2) a procedure for exosome isolation with several gradient alternatives; and (3) post-isolation procedures for estimating the purity and yield of the exosomal fraction.

Place, publisher, year, edition, pages
Wiley-Blackwell, 2016. Vol. 2016, p. 14.42.1-14.42.21
Keywords [en]
density ultracentrifugation, exosomes, Extracellular vesicles, flow cytometry, negative contrast staining, Organ explant culture, sucrose gradient, TEM
National Category
Immunology in the medical area Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-200033DOI: 10.1002/cpim.17PubMedID: 27801511Scopus ID: 2-s2.0-85018275366OAI: oai:DiVA.org:umu-200033DiVA, id: diva2:1702044
Available from: 2022-10-10 Created: 2022-10-10 Last updated: 2022-10-10Bibliographically approved

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Mincheva-Nilsson, LuciaBaranov, VladimirNagaeva, OlgaDehlin, Eva

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  • apa
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