A novel human gastric primary cell culture system for modelling Helicobacter pylori infection in vitro

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Abstract

Background and aims

Helicobacter pylori is the causative agent of gastric diseases and the main risk factor in the development of gastric adenocarcinoma. In vitro studies with this bacterial pathogen largely rely on the use of transformed cell lines as infection model. However, this approach is intrinsically artificial and especially inappropriate when it comes to investigating the mechanisms of cancerogenesis. Moreover, common cell lines are often defective in crucial signalling pathways relevant to infection and cancer. A long-lived primary cell system would be preferable in order to better approximate the human in vivo situation.

Methods

Gastric glands were isolated from healthy human stomach tissue and grown in Matrigel containing media supplemented with various growth factors, developmental regulators and apoptosis inhibitors to generate long-lasting normal epithelial cell cultures.

Results

Culture conditions were developed which support the formation and quasi-indefinite growth of three dimensional (3D) spheroids derived from various sites of the human stomach. Spheroids could be differentiated to gastric organoids after withdrawal of Wnt3A and R-spondin1 from the medium. The 3D cultures exhibit typical morphological features of human stomach tissue. Transfer of sheared spheroids into 2D culture led to the formation of dense planar cultures of polarised epithelial cells serving as a suitable in vitro model of H. pylori infection.

Conclusions

A robust and quasi-immortal 3D organoid model has been established, which is considered instrumental for future research aimed to understand the underlying mechanisms of infection, mucosal immunity and cancer of the human stomach.

Related collections

Most cited references 35

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We previously estimated that 660,000 cases of cancer in the year 2008 were attributable to the bacterium Helicobacter pylori (H. pylori), corresponding to 5.2% of the 12.7 million total cancer cases that occurred worldwide. In recent years, evidence has accumulated that immunoblot (western blot) is more sensitive for detection of anti-H. pylori antibodies than ELISA, the detection method used in our previous analysis. The purpose of this short report is to update the attributable fraction (AF) estimate for H. pylori after briefly reviewing new evidence, and to reassess the global burden of cancer attributable to H. pylori. We therefore reviewed the literature for studies comparing the risk of developing non-cardia gastric cancer (NCGC) in cases and controls, using both ELISA and multiple antigen immunoblot for detection of H. pylori. The results from prospective studies were combined, and the new pooled estimates were applied to the calculation of the AF for H. pylori in NCGC, then to the burden of infection-related cancers worldwide. Using the immunoblot-based data, the worldwide AF for H. pylori in NCGC increased from 74.7% to 89.0%. This implies approximately 120,000 additional cases of NCGC attributable to H. pylori infection for a total of around 780,000 cases (6.2% instead of 5.2% of all cancers). These updated estimates reinforce the role of H. pylori as a major cause of cancer.

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Author and article information

Journal

Journal ID (nlm-ta): Gut

Journal ID (iso-abbrev): Gut

Journal ID (hwp): gutjnl

Journal ID (publisher-id): gut

Title: Gut

Publisher: BMJ Publishing Group (BMA House, Tavistock Square, London, WC1H 9JR )

ISSN (Print): 0017-5749

ISSN (Electronic): 1468-3288

Publication date (Print): February 2016

Publication date (Electronic): 24 December 2014

Volume: 65

Issue: 2

Pages: 202-213

Affiliations

[1 ]Department of Molecular Biology, Max Planck Institute for Infection Biology , Berlin, Germany

[2 ]Clinics for General, Visceral and Transplant Surgery, Charité University Medicine , Berlin, Germany

[3 ]Center of Bariatric and Metabolic Surgery, Charité University Medicine , Berlin, Germany

[4 ]Hubrecht Institute/KNAW and University Medical Centre Utrecht , Utrecht, The Netherlands

Author notes

[Correspondence to ] Professor Dr Thomas F Meyer, Department of Molecular Biology, Max Planck Institute for Infection Biology, Charitéplatz 1, Berlin 10117, Germany; meyer@ 123456mpiib-berlin.mpg.de

Author information

Thomas F Meyer http://orcid.org/0000-0002-6120-8679

Article

Publisher ID: gutjnl-2014-307949

DOI: 10.1136/gutjnl-2014-307949

PMC ID: 4752654

PubMed ID: 25539675

SO-VID: d126e77d-012a-44d6-b104-a5c42de75362

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License:

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