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A human intestinal M-cell-like model for investigating particle, antigen and microorganism translocation
Catholic Univ Louvain, Louvain Drug Res Inst, Dept Adv Drug Delivery & Biomat, Brussels, Belgium..
Univ Coll Dublin, Sch Vet Med, Vet Biosci Sect, Dublin, Ireland.;Univ Coll Dublin, Conway Inst, Dublin, Ireland..
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
Catholic Univ Louvain, Louvain Drug Res Inst, Dept Adv Drug Delivery & Biomat, Brussels, Belgium..
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2017 (English)In: Nature Protocols, ISSN 1754-2189, E-ISSN 1750-2799, Vol. 12, no 7, 1387-1399 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP , 2017. Vol. 12, no 7, 1387-1399 p.
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-328973DOI: 10.1038/nprot.2017.041ISI: 000403344900005PubMedID: 28617450OAI: oai:DiVA.org:uu-328973DiVA: diva2:1140570
Funder
EU, FP7, Seventh Framework Programme, 281035
Available from: 2017-09-12 Created: 2017-09-12 Last updated: 2017-09-12Bibliographically approved

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Artursson, Per

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