A human intestinal M-cell-like model for investigating particle, antigen and microorganism translocation

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Title: A human intestinal M-cell-like model for investigating particle, antigen and microorganism translocation
Authors: Beloqui, Ana
Brayden, David James
Artursson, Per
Préat, Véronique
des Rieux, Anne
Permanent link: http://hdl.handle.net/10197/9138
Date: 15-Jun-2017
Abstract: The specialized microfold cells (M cells) in the follicle-associated epithelium (FAE) of intestinal Peyer's patches serve as antigen-sampling cells of the intestinal innate immune system. Unlike 'classical' enterocytes, they are able to translocate diverse particulates without digesting them. They act as pathways for microorganism invasion and mediate food tolerance by transcellular transport of intestinal microbiota and antigens. Their ability to transcytose intact particles can be used to develop oral drug delivery and oral immunization strategies. This protocol describes a reproducible and versatile human M-cell-like in vitro model. This model can be exploited to evaluate M-cell transport of microparticles and nanoparticles for protein, drug or vaccine delivery and to study bacterial adherence and translocation across M cells. The inverted in vitro M-cell model consists of three main steps. First, Caco-2 cells are seeded at the apical side of the inserts. Second, the inserts are inverted and B lymphocytes are seeded at the basolateral side of the inserts. Third, the conversion to M cells is assessed. Although various M-cell culture systems exist, this model provides several advantages over the rest: (i) it is based on coculture with well-established differentiated human cell lines; (ii) it is reproducible under the conditions described herein; (iii) it can be easily mastered; and (iv) it does not require the isolation of primary cells or the use of animals. The protocol requires skills in cell culture and microscopy analysis. The model is obtained after 3 weeks, and transport experiments across the differentiated model can be carried out over periods of up to 10 h..
Funding Details: European Commission - Seventh Framework Programme (FP7)
Type of material: Journal Article
Publisher: Springer Nature
Copyright (published version): 2017 Macmillan Publishers/Springer Nature
Keywords: M cellsPeyer's patchesOral vaccinesNanoparticles
DOI: 10.1038/nprot.2017.041
Language: en
Status of Item: Peer reviewed
Appears in Collections:Veterinary Medicine Research Collection

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