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Isolation and Functional Studies of Human Fetal Gastric Epithelium in Primary Culture

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Human Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 806))

Abstract

Our understanding of gastric epithelial physiology in man is limited by the absence of normal or appropriate cancer cell lines that could serve as an in vitro model. Research mostly relied on primary culture of gastric epithelial cells of animal species, enriched with surface mucous cells, and devoid of glandular zymogenic chief cells. We successfully applied a new nonenzymatic procedure using Matrisperse Cell Recovery Solution to dissociate the entire epithelium from human fetal stomach. Cultures were generated by seeding multicellular aggregates prepared by mechanical fragmentation. We further demonstrate that this simple and convenient technique allows for the maintenance of heterogenous gastric epithelial primary cultures on plastic without a biological matrix as well as the persistence of viable chief cells able to synthesize and secrete gastric digestive enzymes, i.e., pepsinogen and gastric lipase. In wounding experiments, epithelial restitution occurred in serum-reduced conditions and was modulated by exogenous agents. This culture system is thus representative of the foveolus-gland axis and offers new perspectives to establish the influence of individual growth factors and extracellular matrix components as well as their combinatory effects on gastric epithelium homeostasis.

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Acknowledgments

This work was supported by Canadian Institutes of Health Research (CIHR) grant MOP-36495 to D.M. J.F.B. and D.M. are members of the FRSQ-funded Centre de Recherche Clinique Étienne-LeBel from the CHUS. J.F.B. is the recipient of the Canadian Research Chair in Intestinal Physiopathology.

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Authors and Affiliations

  1. CIHR Team on Digestive Epithelium, Département d’anatomie et de biologie cellulaire, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, Sherbrooke, QC, Canada

    Pierre Chailler, Jean-François Beaulieu & Daniel Ménard

Authors
  1. Pierre Chailler
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  2. Jean-François Beaulieu
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  3. Daniel Ménard
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Corresponding author

Correspondence to Daniel Ménard .

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Editors and Affiliations

  1. Institute of Liver Studies, King’s College Hospital, Denmark Hill, London, SE5 9RS, United Kingdom

    Ragai R. Mitry

  2. School of Medicine, Inst. Liver Studies, King's College London, Bessemer Rd., London, SE5 9RS, United Kingdom

    Robin D. Hughes

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Chailler, P., Beaulieu, JF., Ménard, D. (2012). Isolation and Functional Studies of Human Fetal Gastric Epithelium in Primary Culture. In: Mitry, R., Hughes, R. (eds) Human Cell Culture Protocols. Methods in Molecular Biology, vol 806. Humana Press. https://doi.org/10.1007/978-1-61779-367-7_10

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  • DOI: https://doi.org/10.1007/978-1-61779-367-7_10

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-366-0

  • Online ISBN: 978-1-61779-367-7

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