Conditionally immortalized gastric epithelial cell lines were established from transgenic @entity35 harboring temperature-sensitive @entity2235 (tsSV40) large T-antigen gene. Gastric mucosal cells and epithelial tissues isolated from the stomach of the transgenic @entity35 were cultured at permissive temperature (33 degrees C), and proliferative cells were cloned by colony formation. Six cell lines (designated as RGE1-01, RGE1-02, RGE1-03, RGE1-21, RGE1-22 and RGE2-01) showing epithelial-like morphology have been established. All cells grew at 33 degrees C, but did not at nonpermissive temperature (39 degrees C). High expression level of large T-antigen in the nuclei was observed at 33 degrees C, whereas the expression level was gradually decreased in a time-dependent manner at 39 degrees C. These results suggest that the temperature-sensitive growth characteristics arise as a result of a function of the tsSV40 large T-antigen. None of the cell lines were transformed as judged by anchorage-independent growth assay. Immunocytochemical findings indicated that all cells expressed epithelial cell markers including cytoskeletal (cytokeratin and actin), basement membrane (laminin and collagen type IV) and junctional complex (ZO-1 and desmoplakin I+II) proteins at 33 degrees C. All cells expressed mRNA of @entity14408 , a pit cell marker. Moreover, transepithelial resistance was observed between apical and basolateral sides in the cells. RGE1-22 cells produced @entity1421 . Levels of mRNA for @entity14408 , transepithelial resistance and @entity1421 were influenced by the nonpermissive temperature. Thus, these conditionally immortalized gastric cell lines which preserve some epithelial cell characteristics will provide a useful in vitro model of gastric epithelium.

Development and characterization of conditionally immortalized gastric epithelial cell lines from transgenic XXXX harboring temperature-sensitive @entity2235 large T-antigen gene.

@entity35 :: ('10116', 'Species') :: ['rats'] @entity1421 :: ('MESH:D015232', 'Chemical') :: ['prostaglandin E(2)'] @entity14408 :: ('25424(Tax:10116)', 'Gene') :: ['cathepsin E'] @entity2235 :: ('10633', 'Species') :: ['simian virus 40']
@entity35 :: (10116,Species) :: ['rats']
