Three-dimensional organotypic co-culture model of intestinal epithelial cells and macrophages to study Salmonella enterica colonization patterns

Jennifer Barrila, Jiseon Yang, Aurélie Crabbé, Shameema F. Sarker, Yulong Liu, C. Mark Ott, Mayra A. Nelman-Gonzalez, Simon J. Clemett, Seth D. Nydam, Rebecca J. Forsyth, Richard R. Davis, Brian E. Crucian, Heather Quiriarte, Kenneth L. Roland, Karen Brenneman, Clarence Sams, Christine Loscher, Cheryl Nickerson

Research output: Contribution to journalArticlepeer-review

29 Scopus citations


Three-dimensional models of human intestinal epithelium mimic the differentiated form and function of parental tissues often not exhibited by two-dimensional monolayers and respond to Salmonella in key ways that reflect in vivo infections. To further enhance the physiological relevance of three-dimensional models to more closely approximate in vivo intestinal microenvironments encountered by Salmonella, we developed and validated a novel three-dimensional co-culture infection model of colonic epithelial cells and macrophages using the NASA Rotating Wall Vessel bioreactor. First, U937 cells were activated upon collagen-coated scaffolds. HT-29 epithelial cells were then added and the three-dimensional model was cultured in the bioreactor until optimal differentiation was reached, as assessed by immunohistochemical profiling and bead uptake assays. The new co-culture model exhibited in vivo-like structural and phenotypic characteristics, including three-dimensional architecture, apical-basolateral polarity, well-formed tight/adherens junctions, mucin, multiple epithelial cell types, and functional macrophages. Phagocytic activity of macrophages was confirmed by uptake of inert, bacteria-sized beads. Contribution of macrophages to infection was assessed by colonization studies of Salmonella pathovars with different host adaptations and disease phenotypes (Typhimurium ST19 strain SL1344 and ST313 strain D23580; Typhi Ty2). In addition, Salmonella were cultured aerobically or microaerobically, recapitulating environments encountered prior to and during intestinal infection, respectively. All Salmonella strains exhibited decreased colonization in co-culture (HT-29-U937) relative to epithelial (HT-29) models, indicating antimicrobial function of macrophages. Interestingly, D23580 exhibited enhanced replication/survival in both models following invasion. Pathovar-specific differences in colonization and intracellular co-localization patterns were observed. These findings emphasize the power of incorporating a series of related three-dimensional models within a study to identify microenvironmental factors important for regulating infection.

Original languageEnglish (US)
Article number10
Journalnpj Microgravity
Issue number1
StatePublished - Dec 1 2017

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Materials Science (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Agricultural and Biological Sciences (miscellaneous)
  • Physics and Astronomy (miscellaneous)
  • Space and Planetary Science


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