title: Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche.
abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell-cell and epithelial-mesenchymal interactions in an environment that is extremely similar to the in vivo environment.
authors.affiliation: Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Nagoya 467-8601, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Katano
authors.forename: Akifumi
authors.initials: A
authors.lastname: Ootani
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Mizoshita
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanida
authors.forename: Hironobu
authors.initials: H
authors.lastname: Tsukamoto
authors.forename: Keiji
authors.initials: K
authors.lastname: Ozeki
authors.forename: Masahide
authors.initials: M
authors.lastname: Ebi
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Mori
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kataoka
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kamiya
authors.forename: Shuji
authors.initials: S
authors.lastname: Toda
authors.forename: Takashi
authors.initials: T
authors.lastname: Joh
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D055271:Mucin 5AC / Q000378:metabolism; D061251:Primary Cell Culture; D055153:Stem Cell Niche
chemical_list: C524679:Muc5ac protein, mouse; D055271:Mucin 5AC