In Vitro Gut Model to Assess Bacterial Adhesion and Internalization

Begin with a Transwell insert containing a co-culture of epithelial and goblet-like cells.

Epithelial cells form tight junctions, while goblet-like cells produce mucus, replicating the in vivo intestinal epithelial barrier.

Take a suspension of the oral bacterial community that has been treated with simulated intestinal fluid to mimic small intestinal digestion.

Replace the media in the upper chamber with the bacterial suspension.

Incubate with shaking to facilitate bacterial adhesion and internalization.

Collect the media from the upper chamber containing non-adherent bacteria for subsequent viability assays.

Add a mucus-dissolving solution and incubate to solubilize the mucus and release the adhered bacteria.

Collect the solution for further analysis of bacterial viability and adhesion.

Wash the cells with buffer and add a detergent to dislodge them.

Collect the cell suspension.

Vortex the suspension to lyse the cells and release the internalized bacteria.

Centrifuge and collect the supernatant and cellular debris separately for further analysis.

After washing and adjusting the cell density of the small intestine digestion according to the text protocol, remove the apical medium from the tra