BACKGROUND/AIMS: The unavailability of human gastric cell lines representative of the normal gastric epithelial function such as polarized monolayer restricts the application of cell culture system in approaching the field of Helicobacter pylori (H. pylori) infected gastric mucosa models. The present investigation aimed at assessing the usefulness of NCI-N87 cell line as an adequate cellular model to study the pathophysiology of human H. pylori infection. METHODS: For the identification of epithelial phenotypes at low magnification, cells were observed on a phase-contrast microscope and confocal microscope. Transepithelial resistance (TER) was measured on NCI-N87 cells seeded on Transwell to identify monolayer polarity two or three times a week after confluency. The IL-8 level was determined by ELISA at 24 hours after the administration of HP60190 and IL-1 alpha on NCI-N87 cells. IL-8 level was compared in both upper and lower well with the control. RESULTS: A monolayer phenotype was observed in NCI-N87 cell lines by using confocal microscope. TER was measured as 400-500 (Omega x cm2) at two or three weeks after cell culture. In NCI-N87 cell lines, IL-8 level was significantly increased after 24 hour compared to control, and was prominent in the lower well. CONCLUSIONS: These results suggest that NCI-N87 cell line may be useful in H. pylori infected gastric mucosa model.
BACKGROUND/AIMS: The unavailability of human gastric cell lines representative of the normal gastric epithelial function such as polarized monolayer restricts the application of cell culture system in approaching the field of Helicobacter pylori (H. pylori) infected gastric mucosa models. The present investigation aimed at assessing the usefulness of NCI-N87 cell line as an adequate cellular model to study the pathophysiology of humanH. pyloriinfection. METHODS: For the identification of epithelial phenotypes at low magnification, cells were observed on a phase-contrast microscope and confocal microscope. Transepithelial resistance (TER) was measured on NCI-N87 cells seeded on Transwell to identify monolayer polarity two or three times a week after confluency. The IL-8 level was determined by ELISA at 24 hours after the administration of HP60190 and IL-1 alpha on NCI-N87 cells. IL-8 level was compared in both upper and lower well with the control. RESULTS: A monolayer phenotype was observed in NCI-N87 cell lines by using confocal microscope. TER was measured as 400-500 (Omega x cm2) at two or three weeks after cell culture. In NCI-N87 cell lines, IL-8 level was significantly increased after 24 hour compared to control, and was prominent in the lower well. CONCLUSIONS: These results suggest that NCI-N87 cell line may be useful in H. pyloriinfected gastric mucosa model.
Authors: Maria Fiorentino; Hua Ding; Thomas G Blanchard; Steven J Czinn; Marcelo B Sztein; Alessio Fasano Journal: Infect Immun Date: 2013-01-07 Impact factor: 3.441
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