This mutant still induced IL8 expression, indicating that the bacterial flagellum is not the major inducer of IL8 in this system ( Supplementary Figure 4E). We then asked whether specific cells in the organoids respond to the

bacteria and used our differentiation protocol to generate gland-type or pit-type organoids, which we subsequently microinjected with H pylori. IL8 expression was substantially higher in gland-type organoids than in pit-type organoids ( Figure 6F). Here, we present a long-term 3-dimensional organoid culture system for primary, untransformed human gastric epithelium as well as human gastric cancer. By using this culture, we provide SB203580 cell line direct evidence for the presence of stem cells in adult human gastric tissue. The cells can be directed

to differentiate into specific lineages of the stomach. The organoids mount an NF-κB–driven inflammatory response to infection and the strength of this response depends on the differentiated cell types in the organoids. The presence of stem cells in the human adult stomach is expected, yet has not been shown previously. The organoids we present here can be grown from fluorescence-activated cell sorter–isolated single cells and generate 4 lineages of the stomach: pit mucous cells, gland mucous cells, chief cells, and enteroendocrine cells. Of the enteroendocrine cells, we identified SST-expressing cells, but not CP-868596 molecular weight corpus-specific ECL cells. We also could not detect parietal cells. We assume the culture conditions were not optimal to allow differentiation into these cell types. Once clonal organoids are established, they expand without apparent limitation (>1 y), defying the Hayflick limit. Thus, the isolated cells can self-renew and are long-lived and multipotent, fulfilling the classic criteria for stem cells. In the intestine, the pathologic activation of the Wnt pathway in cancer represents a deregulation

of the controlled activation necessary for normal stem cell–driven tissue homeostasis.25 In the stomach, the role of the Wnt pathway is less clear. Up to 30% of gastric tumors are found to carry an activated Wnt pathway,26 and 27 whereas mutations in the Wnt pathway drive tumorigenesis in the mouse.4 and 28 Two of the known stem Verteporfin price cell markers in the mouse stomach, Troy and Lgr5, are Wnt target genes.4 and 11 Here, we provide additional evidence for the importance of the Wnt pathway in human gastric epithelium. First, establishment and growth of human gastric organoids depends on Wnt and R-Spondin1. Second, on withdrawal of Wnt, organoids differentiate into pit lineage cultures. In the intestine, the Wnt-secreting Paneth cells provide the niche for stem cells17 and competition for niche space determines the fate of the stem cell daughter cells.5 and 6 It seems likely that there is a Wnt source at the bottom of gastric glands and that the migration of daughter cells upward toward the gastric surface directs the differentiation into the pit lineage.