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Imaging flow cytometry facilitates multiparametric characterization of extracellular vesicles in malignant brain tumours
| Content Provider | Scilit |
|---|---|
| Author | Ricklefs, Franz L. Maire, Cecile L. Reimer, Rudolph Dührsen, Lasse Kolbe, Katharina Holz, Mareike Schneider, Enja Rissiek, Anne Babayan, Anna Hille, Claudia Pantel, Klaus Krasemann, Susanne Glatzel, Markus Heiland, Dieter Henrik Flitsch, Jörg Martens, Tobias Schmidt, Nils Ole Peine, Sven Breakefield, Xandra O. Lawler, Sean Chiocca, E. Antonio. Fehse, Boris Giebel, Bernd Görgens, André Westphal, Manfred Lamszus, Katrin |
| Copyright Year | 2019 |
| Description | Cells release heterogeneous nano-sized vesicles either as exosomes, being derived from endosomal compartments, or through budding from the plasma membrane as so-called microvesicles, commonly referred to as extracellular vesicles (EVs). EVs are known for their important roles in mammalian physiology and disease pathogenesis and provide a potential biomarker source in cancer patients. EVs are generally often analysed in bulk using Western blotting or by bead-based flow-cytometry or, with limited parameters, through nanoparticle tracking analysis. Due to their small size, single EV analysis is technically highly challenging. Here we demonstrate imaging flow cytometry (IFCM) to be a robust, multiparametric technique that allows analysis of single EVs and the discrimination of distinct EV subpopulations. We used IFCM to analyse the tetraspanin (CD9, CD63, CD81) surface profiles on EVs from human and murine cell cultures as well as plasma samples. The presence of EV subpopulations with specific tetraspanin profiles suggests that EV-mediated cellular responses are tightly regulated and dependent on cell environment. We further demonstrate that EVs with double positive tetraspanin expression $(CD63^{+}/CD81^{+}$) are enriched in cancer cell lines and patient plasma samples. In addition, we used IFCM to detect tumour-specific GFP-labelled EVs in the blood of mice bearing syngeneic intracerebral gliomas, indicating that this technique allows unprecedented disease modelling. In summary, our study highlights the heterogeneous and adaptable nature of EVs according to their marker profile and demonstrates that IFCM facilitates multiparametric phenotyping of EVs not only in vitro but also in patient plasma at a single EV level, with the potential for future functional studies and clinically relevant applications. Abbreviation: EDTA = ethylenediamine tetraacetic acid |
| Related Links | https://www.tandfonline.com/doi/pdf/10.1080/20013078.2019.1588555?needAccess=true https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6442086/pdf |
| ISSN | 20013078 |
| e-ISSN | 20013078 |
| DOI | 10.1080/20013078.2019.1588555 |
| Journal | Journal of Extracellular Vesicles |
| Issue Number | 1 |
| Volume Number | 8 |
| Language | English |
| Publisher | Informa UK Limited |
| Publisher Date | 2019-12-01 |
| Access Restriction | Open |
| Subject Keyword | Pathology Imaging Flow Cytometry Extracellular Vesicle Biomarker Glioma Tetraspanin |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Histology |
