The gastric pit-gland unit is a highly dynamic and compartimentalized structure which assumes important key functions such as acid secretion, digestion of dietary proteins and triglycerides, protection, and epithelial restitution following injury. However, in vitro models representative of the intact gastric epithelium are still lacking. The current study was undertaken to investigate the possibility of generating such primary cultures from human fetal stomach. The use of Matrisperse, a nonenzymatic solution, allowed complete dissociation of the epithelial layer and the maintenance for at least 7 days of all gastric epithelial cell types in primary culture on plastic. Indirect immunofluorescence and Western blot analyses confirmed the purity of epithelial cultures, composed of 60% mucus-secreting cells, 25% zymogenic chief cells, 5% parietal cells, and a small proportion of mitotic precursors. Their functionality was demonstrated by the presence of zonulae occludens and adherens at cell to cell contacts, [(3)H]thymidine incorporation, Periodic acid Schiff staining, and expression of growth factor receptors (EGF/TGFalpha, IGF1, HGF, KGF), gastric H(+)/K(+)-ATPase, pepsinogen (Pg5), and human gastric lipase (HGL). Chief cells were able to produce and secrete both Pg5 and HGL and to respond to EGF treatment. In conclusion, we developed a new primary culture system of human gastric epithelium characterized for the first time by the absence of added matrix and the maintenance of functional chief cells. It represents an experimental breakthrough that will serve applications in investigating the actions of hormones, mesenchymal growth factors, and basement membrane proteins on human gastric functions in vitro.
Copyright 1999 Academic Press.