A novel in vitro infection model of Helicobacter pylori using mucin-producing murine gastric surface mucous cells.
BACKGROUND
Helicobacter pylori is found within the gastric surface mucous gel layer and in the epithelial surface. Gastric cancer cells have been used in experimental H. pylori infection in vitro, although cancer cells have some abnormalities in cellular properties. The aim of this study was to develop an in vitro H. pylori infection model using normal gastric surface cells that produce gastric mucin.


METHODS
Normal murine gastric surface mucous cells (GSM06) were cultured by the liquid interface method using a serum-free medium and a collagen gel containing a fibroblast cell line (L929) and infected with H. pylori. Infection by H. pylori was assessed by enumerating the colony-forming units (CFU) of H. pylori adhered to GSM06 cells and by transmission electron microscopy. The production of mucin was determined by a lectin binding assay, sugar analysis, and MUC5AC gene expression.


RESULTS
GSM06 cells cultured under these conditions produced mucin containing N-acetylgalactosamine and MUC5AC as the core protein. Significantly higher numbers of H. pylori adhered to GSM06 cells under mucin-producing conditions than under nonproducing conditions. Microscopic observation showed a filamentous structure resembling a type IV secretion system apparatus formed between the surface of GSM06 cells and H. pylori.


CONCLUSIONS
This study demonstrates a novel in vitro H. pylori infection model using mucin-producing murine GSM06 cells for early stages of infection.
Graduate School of Pharmaceutical Sciences, Kitasato University, Minato-ku, Tokyo, Japan.
Tetsufumi
T
Takahashi
Tsukasa
T
Matsumoto
Masahiko
M
Nakamura
Hidenori
H
Matsui
Hiroaki
H
Kiyohara
Chihiro
C
Sasakawa
Haruki
H
Yamada
D000116:Acetylgalactosamine / Q000032:analysis; D000818:Animals; D001422:Bacterial Adhesion; D002460:Cell Line; D015169:Colony Count, Microbial; D010861:Fimbriae, Bacterial / Q000648:ultrastructure; D005752:Gastric Mucins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005753:Gastric Mucosa / Q000378:metabolism* / Q000382:microbiology* / Q000648:ultrastructure; D015870:Gene Expression; D016480:Helicobacter pylori / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008854:Microscopy, Electron; D055271:Mucin 5AC; D009077:Mucins / Q000032:analysis / Q000235:genetics; D021381:Protein Transport
D005752:Gastric Mucins; C524679:Muc5ac protein, mouse; D055271:Mucin 5AC; D009077:Mucins; D000116:Acetylgalactosamine