Original scientific paper
Characterization of a Cell-culturing System for the Study of Contact-independent Extracellular Vesicle Communication
Anne Louise Schacht Revenfeld
; Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
Evo Kristina Lindersson Søndergaard
; Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
Allan Stensballe
; Department of Health Science and Technology, Laboratory for Medical Mass Spectrometry Fredrik Bajersvej, Aalborg University ,Aalborg, Denmark
Rikke Bæk
; Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
Malene Møller Jørgensen
; Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
Kim Varming
; Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
Abstract
Appropriate and well-documented in vitro cell-culturing systems are necessary to study the activity and biological function of extracellular vesicles (EVs). The aim of this study was to describe an experimental system, in which dynamic, vesicle-based cell communication can be investigated. A commercially available cell-culturing system was applied to study contact-independent cell communication, which separated two cell populations using a membrane with a pore size of 0.4 μm. The EV exchange characteristics between the two compartments in the culture set-up was preliminarily investigated in a cell-free set-up, and analysed using the Extracellular Vesicle (EV) Array and Nanoparticle Tracking Analysis. The application of the cell-culturing set-up was demonstrated using co-cultures of human primary cells. The effects of the relative placement of the two cell populations on the phenotype of EVs found in the cell supernatant were investigated. The results indicate that this placement can be important for the biological hypothesis that is being investigated. These observations are relevant for short (<24h) as well as long (several days) studies of vesicle-based cell communication. Moreover, the introduced cell-culturing set-up and analytical strategy can be used to study contact-independent vesicle communication in a reproducible manner.
Keywords
Extracellular Vesicles; Cell Communication; Contact-independent; EV Array; Phenotype; Transwell
Hrčak ID:
161461
URI
Publication date:
1.1.2016.
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