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Cancer Research (Subject) / V.10 Cancer exosomes (Lesson)
- Definition of EVs Extracellular vesicles (EV´s) are particles, naturally released from the cell, delimited by a lipid bilayer and unable to replicate.
- Definition of Exosomes Exosomes are EVs that are 30–150 nm in diameter. Their origin is the multivesicular endosome pathway.
- Name sources of exosomes. • Blood plasma and serum• Urine• Saliva• Cerebrospinal fluid (CSF)• Breast milk• Lymph• stool• semen• bronchial fluid• amniotic fluid• synovial fluid• gastric acid• tears • Cell conditioned media from in vitro human and mouse cell lines• Plasma or serum derived from pre-clinical animal model
- Name EVs isolation methods -Ultracentrifugation + density gradient -precipitation -ultrafiltration -size exclusion chromatography -Immunoaffinity Microfluids-based methods -On-chip centrifugation -Acoustophoresis -Filtration -Viscoelastic flow -Immunoaffinity
- Name EV characterization methods -Nanoparticle Tracking Analysis (NTA) -Particles to protein ratio -Transmission electron microscopy and immunogold imaging -Flow cytometry -Western Blotting and ELISA
- Process to visualize EV dsDNA location 1) cells cultured with EdU (5-ethynyl-2'-deoxyuridine ) --> cancer cell line 2) EVs with EdU labeled DNA collected --> transfer to recipient cells 3) cells co-cultured with EVs 4) Fixation, permeabilisation, DAPI staining and confocal microscopy to visualize EV dsDNA location OR Co-localization staining / inhibitors
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