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

Physiological Reports

Yong-Guo Zhang²  Shaoping Wu²  Yinglin Xia¹ 

[1] Department of Biostatistics and Computational Biology, University of Rochester, Rochester, New York;Department of Biochemistry, Rush University, Chicago, Illinois

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

DOI  :  10.14814/phy2.12147

来源: Wiley

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.