Improved in vitro model systems for gastrointestinal infection by choice of cell line, pH, microaerobic conditions, and optimization of culture conditions

Linden, Sara K., Driessen, Kim M. and McGuckin, Michael A. (2007) Improved in vitro model systems for gastrointestinal infection by choice of cell line, pH, microaerobic conditions, and optimization of culture conditions. Helicobacter, 12 4: 341-353. doi:10.1111/j.1523-5378.2007.00509.x


Author Linden, Sara K.
Driessen, Kim M.
McGuckin, Michael A.
Title Improved in vitro model systems for gastrointestinal infection by choice of cell line, pH, microaerobic conditions, and optimization of culture conditions
Journal name Helicobacter   Check publisher's open access policy
ISSN 1083-4389
1523-5378
Publication date 2007-08-01
Sub-type Article (original research)
DOI 10.1111/j.1523-5378.2007.00509.x
Volume 12
Issue 4
Start page 341
End page 353
Total pages 13
Editor D. Y. Graham
Place of publication United Kingdom
Publisher Wiley-Blackwell Publishing
Language eng
Subject 270102 Cell Metabolism
C1
730101 Infectious diseases
Formatted abstract
Background: Commonly used in vitro infection cultures do not mimic the human gastrointestinal tract with regard to pH and microaerobic conditions. Furthermore, despite the importance of mucin–Helicobacter interactions, the cell lines used have not been selected for appropriate mucin expression. To make in vitro studies more applicable to human disease, we have developed coculture methods taking these factors into account.

Materials and methods: Nine human gastrointestinal epithelial cell lines (MKN1, MKN7, MKN28, MKN45, KATO3, HFE145, PCAA/C11 Caco-2, and LS513) were investigated. Expression and glycosylation of mucins (MUC1, 2, 3, 4, 5AC, 5B, 6, 12, 13, and 16) were determined by immunohistochemistry. We analyzed the effect of microaerobic conditions and acidic pH on cell proliferation, viability, and apoptosis.

Results: Microaerobic culture, which is more physiological for the bacteria, did not adversely affect mammalian cell viability, proliferation, or induce apoptosis The cell lines varied in mucin expression, with MKN7 and MKN45 being most similar to gastric mucosa and Caco-2 and LS513 to intestinal mucosa, although none exactly matched normal mucosa. However, changes in culture conditions did not cause major changes in the mucin expression within cell lines.

Conclusions: Culture conditions mimicking the natural environment and allowing the bacterial cells to thrive had no effect on cell viability or apoptosis, and very little influence on mucin expression of human gastrointestinal cells. Thus, it is feasible, using the simple methods we present here, to substantially improve bacterial–mammalian cell in vitro coculture studies to make them more reflective of human infection.
Keyword Mucin
microaerobic culture
bacterial–mammalian co-culture
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2008 Higher Education Research Data Collection
School of Medicine Publications
 
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Created: Wed, 30 Apr 2008, 00:00:56 EST by Maree Knight on behalf of Faculty Of Health Sciences