Gastroenterology

Gastroenterology

Volume 148, Issue 1, January 2015, Pages 126-136.e6
Gastroenterology

Original Research
Full Report: Basic and Translational—Alimentary Tract
In Vitro Expansion of Human Gastric Epithelial Stem Cells and Their Responses to Bacterial Infection

https://doi.org/10.1053/j.gastro.2014.09.042Get rights and content
Under a Creative Commons license
open access

Background & Aims

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.

Keywords

Stomach Cancer
Gastric Epithelium
Primary Cells
Tissue Engineering

Abbreviations used in this paper

cagPAI
cytotoxicity-associated gene pathogenicity island
CDX
caudal-type homeobox
DMEM
Dulbecco’s modified Eagle medium
EGF
epidermal growth factor
ENRWFG
EGF, noggin, R-spondin1, Wnt, FGF10, and gastrin
FGF
fibroblast growth factor
GSK3β
glykogensynthase-Kinase 3 beta
IGF
insulin-like growth factor
LPS
lipopolysaccharide
MOI
multiplicity of infection
MUC
mucin
ODN
oligodeoxynucleotide
PCR
polymerase chain reaction
PGC
pepsinogen
PGE
prostaglandin E
SST
somatostatin
TFF
trefoil factor
TGF
transforming growth factor
TNF
tumor necrosis factor

Cited by (0)

Conflicts of interest These authors disclose the following: Hans Clevers and Meritxell Huch hold patents for organoid culture. The remaining authors disclose no conflicts.

Funding Supported by an EU Marie Curie Fellowship (EU/300686-InfO to S.B. and EU/236954-ICSC-Lgr5 to M.H.), an EU FP7 grant EU/232814-StemCellMark (M.v.d.W. and R.V.), a National Institutes of Health/Massachusetts Institute of Technology subaward 5710002735 (H.B.), and a Research Prize from the United European Gastroenterology Foundation (H.C.).

Author names in bold designate shared co-first authors.