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

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.

A Primary Human Macrophage-Enteroid Co-Culture Model to Investigate Mucosal Gut Physiology and Host-Pathogen Interactions

“…Enteroids and colonoids are reliable human models for the study of viral and bacterial pathogenesis 14,20,21,41,42 . In the present study, we increased the cellular complexity by integrating macrophages to enteroid monolayers with the purpose of interrogating host-pathogen interactions and innate immune response.…”

Translational applications of adult stem cell-derived organoids

“…Co-cultures of mouse small intestinal organoids with Escherichia coli have been used to investigate the effects of antimicrobial products on the dynamics and function of Paneth cells . Moreover, human intestinal organoids were exploited to functionally characterize several cholera toxin inhibitors (Zomervan Ommen et al, 2016), while two recent studies employed mouse intestinal organoids to model Salmonella infections (Wilson et al, 2015;Zhang et al, 2014). In addition to an NFκB-induced inflammatory response, the authors observed disruption of epithelial tight junctions and downregulation of ISC markers Lgr5 and Bmi1 in Salmonella-infected organoids (Zhang et al, 2014).…”

“…Moreover, human intestinal organoids were exploited to functionally characterize several cholera toxin inhibitors (Zomervan Ommen et al, 2016), while two recent studies employed mouse intestinal organoids to model Salmonella infections (Wilson et al, 2015;Zhang et al, 2014). In addition to an NFκB-induced inflammatory response, the authors observed disruption of epithelial tight junctions and downregulation of ISC markers Lgr5 and Bmi1 in Salmonella-infected organoids (Zhang et al, 2014). Neefjes and colleagues recently exploited co-cultures of adult stem cell-derived gallbladder organoids with Salmonella enterica to determine the effects of infectious pathogens on the development of cancer (Scanu et al, 2015).…”

Using 3D Organoid Cultures to Model Intestinal Physiology and Colorectal Cancer

“…While this model has proven useful for studies of barrier function, transcytosis, and cell polarity, it also allows ready infection and detection of attaching and effacing lesions associated with infection [55, 56]. Conversely, intraluminal injection methods have been used to directly inoculate into the enteroid lumen, although this is a much more time consuming model and not amenable to high-throughput applications [57–59]. …”