Foveolar differentiation of mouse gastric mucosa in vitro.
We report a novel method that allows the culture of highly differentiated gastric surface mucous cells. Isolated mouse gastric epithelial cells and fibroblasts were co-cultured in a three-dimensional collagen gel system, and the reconstructed mucosal surface treated with an air-liquid interface. Cultured cells were examined by histology, immunohistochemistry, and electron microscopy. Isolated epithelial cells were positive for MUC5AC, and showed immature mucous cell features (pre-pit cell stage) on cell-free collagen gel. However, when given fibroblastic support, the epithelial cells differentiated into mature surface mucous cells (pit cell stage), and showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm. In the fine structure, the cells showed junctional complexes, basal lamina, and glycogen and secretary granules. Further treatment by the air-liquid interface environment modified the differentiated state of the pit cells (pit top cell stage); resulting in the expression of cathepsin E, the disappearance of glycogen granules and the apical accumulation of secretory granules along with an increase in apoptotic cells. This culture model should provide a useful tool for studying gastric epithelial cell biology and various diseases of the gastric mucosa.
Ootani
Akifumi
A
Department of Pathology, Saga Medical School, Saga, Japan. ohtani@post.saga-med.ac.jp
Toda
Shuji
S
Fujimoto
Kazuma
K
Sugihara
Hajime
H
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D005347:Fibroblasts / Q000166:cytology; D005753:Gastric Mucosa / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D055271:Mucin 5AC; D009077:Mucins / Q000032:analysis
C524679:Muc5ac protein, mouse; D055271:Mucin 5AC; D009077:Mucins; D003094:Collagen