RRC ID 43067
Author Lindén SK, Driessen KM, McGuckin MA.
Title Improved in vitro model systems for gastrointestinal infection by choice of cell line, pH, microaerobic conditions, and optimization of culture conditions.
Journal Helicobacter
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.
Volume 12(4)
Pages 341-53
Published 2007-8-1
DOI 10.1111/j.1523-5378.2007.00509.x
PMID 17669108
MeSH Aerobiosis Apoptosis Caco-2 Cells Cell Culture Techniques / methods Cell Line Cell Survival Cells, Cultured Epithelial Cells / cytology Epithelial Cells / metabolism Epithelial Cells / microbiology* Gastric Mucins / metabolism Gastric Mucosa / cytology Gastric Mucosa / metabolism Gastric Mucosa / microbiology Gastrointestinal Diseases / microbiology Helicobacter Infections / microbiology Helicobacter pylori / growth & development* Humans Hydrogen-Ion Concentration Immunohistochemistry
IF 4.0
Times Cited 31
Human and Animal Cells MKN1(RCB1003) MKN7(RCB0999)