Building a Thick Mucus Hydrogel Layer to Improve the Physiological Relevance of In Vitro Primary Colonic Epithelial Models

摘要

( ) Culture schematic. ( ) Dose-dependent water secretion of VIP at 24 hours. ( ) Immunofluorescence of monolayers differentiated under submerged and ALI conditions in the presence of VIP. Green: anti-Muc2; blue: Hoechst 33342. ( ) Removal of hydrated mucus with forceps. ( ) Side-view confocal micrograph showing tissues with mucus separating bacteria from epithelial cells at 0 and 6 days, respectively. Green: green fluorescent protein (GFP)–expressing ; blue: Hoechst 33342. ( ) Plot of mucus thickness vs duration of ALI. ( ) Representative side-view confocal micrograph showing 1 μm red fluorescent beads separated from epithelial cells by mucus. ( ) Immunofluorescence staining of paraffin-embedded, sectioned monolayers. DM, differentiation medium.