title: Studying extracellular vesicle transfer by a Cre-loxP method.
abstract: Extracellular vesicle (EV) transfer is increasingly recognized as an important mode of intercellular communication by transferring a wide variety of biomolecules between cells. The characterization of in vitro- or ex vivo-isolated EVs has considerably contributed to the understanding of biological functions of EV transfer. However, the study of EV release and uptake in an in vivo setting has remained challenging, because cells that take up EVs could not be discriminated from cells that do not take up EVs. Recently, a technique based on the Cre-loxP system was developed to fluorescently mark Cre-reporter cells that take up EVs released by Cre recombinase-expressing cells in various in vitro and in vivo settings. Here we describe a detailed protocol for the generation of Cre(+) cells and reporter(+) cells, which takes ∼ 6 weeks, and subsequent assays with these lines to study functional EV transfer in in vitro and in vivo (mouse) settings, which take up to ∼ 2 months.
authors.affiliation: Cancer Genomics Netherlands, Hubrecht Institute-Royal Netherlands Academy of Sciences (KNAW) and University Medical Centre Utrecht, Utrecht, the Netherlands.
authors.forename: Anoek
authors.initials: A
authors.lastname: Zomer
authors.affiliation: Cancer Genomics Netherlands, Hubrecht Institute-Royal Netherlands Academy of Sciences (KNAW) and University Medical Centre Utrecht, Utrecht, the Netherlands.
authors.forename: Sander Christiaan
authors.initials: SC
authors.lastname: Steenbeek
authors.affiliation: Cancer Genomics Netherlands, Hubrecht Institute-Royal Netherlands Academy of Sciences (KNAW) and University Medical Centre Utrecht, Utrecht, the Netherlands.
authors.forename: Carrie
authors.initials: C
authors.lastname: Maynard
authors.affiliation: Cancer Genomics Netherlands, Hubrecht Institute-Royal Netherlands Academy of Sciences (KNAW) and University Medical Centre Utrecht, Utrecht, the Netherlands.
authors.forename: Jacco
authors.initials: J
authors.lastname: van Rheenen
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D001692:Biological Transport; D000067128:Extracellular Vesicles / Q000378:metabolism*; D005260:Female; D005821:Genetic Techniques; D057809:HEK293 Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D051379:Mice
chemical_list: C045073:Cre recombinase; D019426:Integrases