Effect of in vitro fermentation derived microbial metabolites on intestinal enteroid function
Abstract
A single‐stage batch in vitro fermentation system has been designed to simulate distal colon microbial activity. This system allows for examination of bacterial function. Outputs include filtered fecal supernatant (FS) that can be used for metabolite identification and examining host response. The goal of this study was to treat human duodenal and colonic enteroids with FS to determine how microbial metabolites alter cell proliferation, differentiation, and expression of other intestinal factors. Three‐dimensional (3D) human duodenal and colonic enteroids were treated with 0, 0.1, 1, 10, 25, and 50% FS or 1μg/mL lipopolysaccharide for 24 h. Cell lysates were used for RNA extraction and mRNA analysis. Visually, increasing FS concentration caused increased browning of enteroids as signs of cell differentiation. At 25 and 50% FS, structural disruption occurred, with fargreater effect at 50% potentially indicating cell death. Expression of LGR5, a stem cell marker, decreased with FS concentration and was lowest at 10% (P 1% FS. Expression of enterocyte and Paneth cells markers were increased by greater than 14‐fold at the 10 and 25% concentrations for alkaline phosphatase (ALPI), and six‐fold at 25% for lysozyme (LYZ), respectively (P PP P PP 8‐fold) in the 25% FS group (P PP P < 0.01). Overall, treatment with FS alters enteroid cell differentiation, and expression of mucin and tight junction protein mRNA, likely similar to a typical intestinal phenotype.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Apr 01, 2019
- Source ID
- 10.1096/fasebj.2019.33.1_supplement.lb543
Entities
People
- Dana M. Van Sambeek
- Jason W. Soares
- Kenneth Racicot
- Laurel A. Doherty
- Sarah C Pearce
Organizations
- United States Army
- United States Department of Defense