title: In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection.
abstract: OBJECTIVE
  We previously established long-term, 3-dimensional culture of organoids from mouse tissues (intestine, stomach, pancreas, and liver) and human intestine and pancreas. Here we describe conditions required for long-term 3-dimensional culture of human gastric stem cells. The technology can be applied to study the epithelial response to infection with Helicobacter pylori.
  
  
  METHODS
  We generated organoids from surgical samples of human gastric corpus. Culture conditions were developed based on those for the mouse gastric and human intestinal systems. We used microinjection to infect the organoids with H pylori. Epithelial responses were measured using microarray and quantitative polymerase chain reaction analyses.
  
  
  RESULTS
  Human gastric cells were expanded indefinitely in 3-dimensional cultures. We cultured cells from healthy gastric tissues, single-sorted stem cells, or tumor tissues. Organoids maintained many characteristics of their respective tissues based on their histology, expression of markers, and euploidy. Organoids from healthy tissue expressed markers of 4 lineages of the stomach and self-organized into gland and pit domains. They could be directed to specifically express either lineages of the gastric gland, or the gastric pit, by addition of nicotinamide and withdrawal of WNT. Although gastric pit lineages had only marginal reactions to bacterial infection, gastric gland lineages mounted a strong inflammatory response.
  
  
  CONCLUSIONS
  We developed a system to culture human gastric organoids. This system can be used to study H pylori infection and other gastric pathologies.
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands. Electronic address: bartfeld@gmx.de.
authors.forename: Sina
authors.initials: S
authors.lastname: Bartfeld
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Tülay
authors.initials: T
authors.lastname: Bayram
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands; Hubrecht Organoid Technology (HUB), Uppsalalaan 8, 3053DC, Utrecht, The Netherlands.
authors.forename: Marc
authors.initials: M
authors.lastname: van de Wetering
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Huch
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Harry
authors.initials: H
authors.lastname: Begthel
authors.affiliation: Division of Cell Biology II, Antoni van Leeuwenhoek Hospital/Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Pekka
authors.initials: P
authors.lastname: Kujala
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands; Hubrecht Organoid Technology (HUB), Uppsalalaan 8, 3053DC, Utrecht, The Netherlands.
authors.forename: Robert
authors.initials: R
authors.lastname: Vries
authors.affiliation: Division of Cell Biology II, Antoni van Leeuwenhoek Hospital/Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Peters
authors.affiliation: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D019070:Cell Lineage; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D005260:Female; D005753:Gastric Mucosa / Q000378:metabolism; D005786:Gene Expression Regulation; D016481:Helicobacter Infections / Q000276:immunology / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D016480:Helicobacter pylori / Q000276:immunology / Q000472:pathogenicity*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009536:Niacinamide / Q000494:pharmacology; D009940:Organoids; D010641:Phenotype; D011003:Ploidies; D013234:Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D013270:Stomach / Q000187:drug effects / Q000276:immunology / Q000382:microbiology* / Q000473:pathology; D013997:Time Factors; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D015415:Biomarkers; D051153:Wnt Proteins; D009536:Niacinamide
keywords: Gastric Epithelium; Primary Cells; Stomach Cancer; Tissue Engineering