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

Sina Bartfeld; Tülay Bayram; Marc van de Wetering; Meritxell Huch; Harry Begthel; Pekka Kujala; Robert Vries; Peter J Peters; Hans Clevers

doi:10.1053/j.gastro.2014.09.042

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

Sina Bartfeld, Tülay Bayram, Marc van de Wetering, Meritxell Huch, Harry Begthel, Pekka Kujala, Robert Vries, Peter J Peters, Hans Clevers

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

612 Citations (Scopus)

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.

Original language	English
Pages (from-to)	126-136.e6
Journal	Gastroenterology
Volume	148
Issue number	1
DOIs	https://doi.org/10.1053/j.gastro.2014.09.042
Publication status	Published - Jan 2015

Keywords

Adult
Aged
Aged, 80 and over
Biological Markers
Cell Culture Techniques
Cell Lineage
Cell Proliferation
Cell Separation
Cells, Cultured
Epithelial Cells
Female
Gene Expression Regulation
Helicobacter Infections
Helicobacter pylori
Humans
Male
Middle Aged
Niacinamide
Organoids
Phenotype
Ploidies
Stem Cells
Stomach
Time Factors
Wnt Proteins

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10.1053/j.gastro.2014.09.042

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title = "In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection",

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 = "Adult, Aged, Aged, 80 and over, Biological Markers, Cell Culture Techniques, Cell Lineage, Cell Proliferation, Cell Separation, Cells, Cultured, Epithelial Cells, Female, Gene Expression Regulation, Helicobacter Infections, Helicobacter pylori, Humans, Male, Middle Aged, Niacinamide, Organoids, Phenotype, Ploidies, Stem Cells, Stomach, Time Factors, Wnt Proteins",

author = "Sina Bartfeld and T{\"u}lay Bayram and {van de Wetering}, Marc and Meritxell Huch and Harry Begthel and Pekka Kujala and Robert Vries and Peters, {Peter J} and Hans Clevers",

year = "2015",

month = jan,

doi = "10.1053/j.gastro.2014.09.042",

language = "English",

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pages = "126--136.e6",

journal = "Gastroenterology",

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T1 - In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection

AU - Bartfeld, Sina

AU - Bayram, Tülay

AU - van de Wetering, Marc

AU - Huch, Meritxell

AU - Begthel, Harry

AU - Kujala, Pekka

AU - Vries, Robert

AU - Peters, Peter J

AU - Clevers, Hans

PY - 2015/1

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N2 - 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.

AB - 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.

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KW - Aged

KW - Aged, 80 and over

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KW - Cell Culture Techniques

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KW - Gene Expression Regulation

KW - Helicobacter Infections

KW - Helicobacter pylori

KW - Humans

KW - Male

KW - Middle Aged

KW - Niacinamide

KW - Organoids

KW - Phenotype

KW - Ploidies

KW - Stem Cells

KW - Stomach

KW - Time Factors

KW - Wnt Proteins

U2 - 10.1053/j.gastro.2014.09.042

DO - 10.1053/j.gastro.2014.09.042

M3 - Article

C2 - 25307862

SN - 0016-5085

VL - 148

SP - 126-136.e6

JO - Gastroenterology

JF - Gastroenterology

IS - 1

ER -

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

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Keywords

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