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Physiol Rep. 2014 Sep 11;2(9). pii: e12147. doi: 10.14814/phy2.12147. Print 2014 Sep 1.

Salmonella-infected crypt-derived intestinal organoid culture system for host-bacterial interactions.

Author information

1
Department of Biochemistry, Rush University, 1735 W. Harrison St., Chicago, Illinois.
2
Department of Biostatistics and Computational Biology, University of Rochester, Rochester, New York.
3
Department of Biochemistry, Rush University, 1735 W. Harrison St., Chicago, Illinois Department of Internal Medicine (GI), Rush University, Chicago, Illinois Department of Microbiology/Immunology, Rush University, Chicago, Illinois.

Abstract

The in vitro analysis of bacterial-epithelial interactions in the intestine has been hampered by a lack of suitable intestinal epithelium culture systems. Here, we report a new experimental model using an organoid culture system to study pathophysiology of bacterial-epithelial interactions post Salmonella infection. Using crypt-derived mouse intestinal organoids, we were able to visualize the invasiveness of Salmonella and the morphologic changes of the organoids. Importantly, we reported bacteria-induced disruption of epithelial tight junctions in the infected organoids. In addition, we showed the inflammatory responses through activation of the NF-κB pathway in the organoids. Moreover, our western blot, PCR, and immunofluorescence data demonstrated that stem cell markers (Lgr5 and Bmi1) were significantly decreased by Salmonella infection (determined using GFP-labeled Lgr5 organoids). For the first time, we created a model system that recapitulated a number of observations from in vivo studies of the Salmonella-infected intestine, including bacterial invasion, altered tight junctions, inflammatory responses, and decreased stem cells. We have demonstrated that the Salmonella-infected organoid culture system is a new experimental model suitable for studying host-bacterial interactions.

KEYWORDS:

Bacteria; Claudin; Lgr5; NF‐κB; ZO‐1; host–bacterial interactions; infection; inflammation; intestinal stem cells; organoid; stem cells; tight junction

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