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  1. Article ; Online: Generation, Characterization, and Count of Fluorescent Extracellular Vesicles.

    Ferrantelli, Flavia / Tirelli, Valentina / Barreca, Valeria / Manfredi, Francesco

    Methods in molecular biology (Clifton, N.J.)

    2022  Volume 2504, Page(s) 207–217

    Abstract: Extracellular vesicles (EVs) are membranous particles released by all cells in the external milieu. Depending on their origin, they are given different names: exosomes are nanovesicles that originate from the endosomal compartment, whereas microvesicles ... ...

    Abstract Extracellular vesicles (EVs) are membranous particles released by all cells in the external milieu. Depending on their origin, they are given different names: exosomes are nanovesicles that originate from the endosomal compartment, whereas microvesicles bud from plasma membrane. Both contain molecules that are crucial for the onset and spreading of different pathologies, from neurodegenerative diseases to cancer, and are considered promising disease markers. On the other hand, EVs are often used as therapeutic tools, and can be engineered to carry drugs and chemicals. This chapter describes a method to produce EVs, mainly exosomes, containing the green fluorescent protein (GFP) linked to an exosome anchoring protein (Nef
    MeSH term(s) Cell-Derived Microparticles/metabolism ; Exosomes/metabolism ; Extracellular Vesicles/metabolism ; Flow Cytometry
    Language English
    Publishing date 2022-04-25
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2341-1_15
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Exploiting Manipulated Small Extracellular Vesicles to Subvert Immunosuppression at the Tumor Microenvironment through Mannose Receptor/CD206 Targeting.

    Fiani, Maria Luisa / Barreca, Valeria / Sargiacomo, Massimo / Ferrantelli, Flavia / Manfredi, Francesco / Federico, Maurizio

    International journal of molecular sciences

    2020  Volume 21, Issue 17

    Abstract: Immunosuppression at tumor microenvironment (TME) is one of the major obstacles to be overcome for an effective therapeutic intervention against solid tumors. Tumor-associated macrophages (TAMs) comprise a sub-population that plays multiple pro-tumoral ... ...

    Abstract Immunosuppression at tumor microenvironment (TME) is one of the major obstacles to be overcome for an effective therapeutic intervention against solid tumors. Tumor-associated macrophages (TAMs) comprise a sub-population that plays multiple pro-tumoral roles in tumor development including general immunosuppression, which can be identified in terms of high expression of mannose receptor (MR or CD206). Immunosuppressive TAMs, like other macrophage sub-populations, display functional plasticity that allows them to be re-programmed to inflammatory macrophages. In order to mitigate immunosuppression at the TME, several efforts are ongoing to effectively re-educate pro-tumoral TAMs. Extracellular vesicles (EVs), released by both normal and tumor cells types, are emerging as key mediators of the cell to cell communication and have been shown to have a role in the modulation of immune responses in the TME. Recent studies demonstrated the enrichment of high mannose glycans on the surface of small EVs (sEVs), a subtype of EVs of endosomal origin of 30-150 nm in diameter. This characteristic renders sEVs an ideal tool for the delivery of therapeutic molecules into MR/CD206-expressing TAMs. In this review, we report the most recent literature data highlighting the critical role of TAMs in tumor development, as well as the experimental evidences that has emerged from the biochemical characterization of sEV membranes. In addition, we propose an original way to target immunosuppressive TAMs at the TME by endogenously engineered sEVs for a new therapeutic approach against solid tumors.
    MeSH term(s) Animals ; Extracellular Vesicles/immunology ; Extracellular Vesicles/metabolism ; Extracellular Vesicles/pathology ; Humans ; Immune Tolerance/immunology ; Lectins, C-Type/metabolism ; Macrophages/immunology ; Macrophages/metabolism ; Macrophages/pathology ; Mannose-Binding Lectins/metabolism ; Neoplasms/immunology ; Neoplasms/metabolism ; Neoplasms/pathology ; Receptors, Cell Surface/metabolism ; Tumor Microenvironment/immunology
    Chemical Substances Lectins, C-Type ; Mannose-Binding Lectins ; Receptors, Cell Surface ; mannose receptor
    Language English
    Publishing date 2020-08-31
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms21176318
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Metabolic labelling of a subpopulation of small extracellular vesicles using a fluorescent palmitic acid analogue.

    Barreca, Valeria / Boussadia, Zaira / Polignano, Deborah / Galli, Lorenzo / Tirelli, Valentina / Sanchez, Massimo / Falchi, Mario / Bertuccini, Lucia / Iosi, Francesca / Tatti, Massimo / Sargiacomo, Massimo / Fiani, Maria Luisa

    Journal of extracellular vesicles

    2023  Volume 12, Issue 12, Page(s) e12392

    Abstract: Exosomes are among the most puzzling vehicles of intercellular communication, but several crucial aspects of their biogenesis remain elusive, primarily due to the difficulty in purifying vesicles with similar sizes and densities. Here we report an ... ...

    Abstract Exosomes are among the most puzzling vehicles of intercellular communication, but several crucial aspects of their biogenesis remain elusive, primarily due to the difficulty in purifying vesicles with similar sizes and densities. Here we report an effective methodology for labelling small extracellular vesicles (sEV) using Bodipy FL C16, a fluorescent palmitic acid analogue. In this study, we present compelling evidence that the fluorescent sEV population derived from Bodipy C16-labelled cells represents a discrete subpopulation of small exosomes following an intracellular pathway. Rapid cellular uptake and metabolism of Bodipy C16 resulted in the incorporation of fluorescent phospholipids into intracellular organelles specifically excluding the plasma membrane and ultimately becoming part of the exosomal membrane. Importantly, our fluorescence labelling method facilitated accurate quantification and characterization of exosomes, overcoming the limitations of nonspecific dye incorporation into heterogeneous vesicle populations. The characterization of Bodipy-labelled exosomes reveals their enrichment in tetraspanin markers, particularly CD63 and CD81, and in minor proportion CD9. Moreover, we employed nanoFACS sorting and electron microscopy to confirm the exosomal nature of Bodipy-labelled vesicles. This innovative metabolic labelling approach, based on the fate of a fatty acid, offers new avenues for investigating exosome biogenesis and functional properties in various physiological and pathological contexts.
    MeSH term(s) Extracellular Vesicles/metabolism ; Palmitic Acid/metabolism ; Exosomes/metabolism ; Biological Transport
    Chemical Substances 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene ; Palmitic Acid (2V16EO95H1)
    Language English
    Publishing date 2023-12-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2683797-3
    ISSN 2001-3078 ; 2001-3078
    ISSN (online) 2001-3078
    ISSN 2001-3078
    DOI 10.1002/jev2.12392
    Database MEDical Literature Analysis and Retrieval System OnLINE

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