Salmonella‐infected crypt‐derived intestinal organoid culture system for host–bacterial interactions

• Published in: Physiological reports Vol. 2; no. 9; pp. e12147 - n/a
• Main Authors: Zhang, Yong‐Guo, Wu, Shaoping, Xia, Yinglin, Sun, Jun
• Format: Journal Article
• Language: English
• Published: United States John Wiley & Sons, Inc 01.09.2014; Wiley Periodicals, Inc
• Subjects: Bacteria; Bacterial infections; Cell culture; Cell division; Claudin; Epithelium; host–bacterial interactions; Immunofluorescence; infection; Infections; Inflammation; intestinal stem cells; Intestine; Invasiveness; Lgr5; Mammals; NF‐κB; organoid; Organoids; Original Research; Physiology; Salmonella; Stem cell transplantation; Stem cells; Studies; tight junction; Tight junctions; ZO‐1; stem cells; infection; NF‐κB; inflammation; Claudin; host–bacterial interactions; intestinal stem cells; Bacteria; Lgr5; ZO‐1; tight junction; organoid
• ISSN: 2051-817X; 2051-817X
• DOI: 10.14814/phy2.12147

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. e12147 Using stem cell‐derived intestinal organoids, we were able to visualize the invasiveness of Salmonella, morphologic changes of the organoids, and showed the inflammatory responses through activating the NF‐κB pathway in the organoids. Moreover, we show that stem cell markers (Lgr5 and Bmi1) were decreased by Salmonella infection. Taken together, the model system we have recreated recapitulates for the first time a number of observations from in vivo studies of Salmonella‐infected gut. Thus, Salmonella‐infected organoid culture system is a new experimental model suitable for studying host–bacterial interactions.