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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.

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Department of Biochemistry, Rush University, 1735 W. Harrison St., Chicago, Illinois.
Department of Biostatistics and Computational Biology, University of Rochester, Rochester, New York.
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.


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.


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

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