Human colonoid monolayers to study interactions between pathogens, commensals, and host intestinal epithelium

Julie In, Jennifer Foulke-Abel, Elizabeth Clarke, Olga Kovbasnjuk

Research output: Contribution to journalArticle

Abstract

Human 3-dimensional (3D) enteroid or colonoid cultures derived from crypt base stem cells are currently the most advanced ex vivo model of the intestinal epithelium. Due to their closed structures and significant supporting extracellular matrix, 3D cultures are not ideal for host-pathogen studies. Enteroids or colonoids can be grown as epithelial monolayers on permeable tissue culture membranes to allow manipulation of both luminal and basolateral cell surfaces and accompanying fluids. This enhanced luminal surface accessibility facilitates modeling bacterial-host epithelial interactions such as the mucus-degrading ability of enterohemorrhagic E. coli (EHEC) on colonic epithelium. A method for 3D culture fragmentation, monolayer seeding, and transepithelial electrical resistance (TER) measurements to monitor the progress towards confluency and differentiation are described. Colonoid monolayer differentiation yields secreted mucus that can be studied by the immunofluorescence or immunoblotting techniques. More generally, enteroid or colonoid monolayers enable a physiologically-relevant platform to evaluate specific cell populations that may be targeted by pathogenic or commensal microbiota.

Original languageEnglish (US)
Article numbere59357
JournalJournal of Visualized Experiments
Volume2019
Issue number146
DOIs
StatePublished - Apr 2019

Keywords

  • Apical and basolateral secretion
  • Apical infection
  • Colonic mucus
  • Host-pathogen interaction
  • Human enteroid or colonoid monolayers
  • Immunology and Infection
  • Intestinal organoid monolayer
  • Issue 146

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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