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.
Original language | English (US) |
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Article number | e12147 |
Journal | Physiological Reports |
Volume | 2 |
Issue number | 9 |
DOIs | |
State | Published - 2014 |
Bibliographical note
Publisher Copyright:© 2014 The Authors.
Keywords
- Bacteria
- Claudin
- Host–bacterial interactions
- Infection
- Inflammation
- Intestinal stem cells
- Lgr5
- NF-κB
- Organoid
- Stem cells
- Tight junction
- ZO-1