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Gastroenterology. 2015 Jan;148(1):126-136.e6. doi: 10.1053/j.gastro.2014.09.042. Epub 2014 Oct 13.

In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection.

Author information

1
Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands. Electronic address: bartfeld@gmx.de.
2
Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, Utrecht, The Netherlands.
3
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.
4
Division of Cell Biology II, Antoni van Leeuwenhoek Hospital/Netherlands Cancer Institute, Amsterdam, The Netherlands.

Abstract

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:

Gastric Epithelium; Primary Cells; Stomach Cancer; Tissue Engineering

PMID:
25307862
PMCID:
PMC4274199
DOI:
10.1053/j.gastro.2014.09.042
[Indexed for MEDLINE]
Free PMC Article
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