Recreating gut-liver axis during NAFLD onset by using a Caco-2/HepG2 co-culture system

Meroni, Marica; Paolini, Erika; Longo, Miriam; Piciotti, Roberto; Tria, Giada; Dongiovanni, Paola

doi:10.20517/mtod.2021.19

Cited by 1 publication

(1 citation statement)

References 36 publications

(44 reference statements)

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“…To create disease models, monolayers were cultured in their respective media supplemented with either 0.2% ethanol, 0.3 mM (2 palmitic acid (PA): 1 oleic acid (OA)) or a combination of 10 µg/ml IL-1β, 10 µg/ml IL-6 and 10 µg/ml TNF-α for 24 hours. These treatments (applied to both the apical and basolateral chambers) have been previous used to stimulate alcoholic lover disease (ALD), 35 Metabolic associated fatty liver disease (MAFLD) 36 and inflammation bowel disease (IBD). 37–39 …”

Section: Methodsmentioning

confidence: 99%

Using a human colonoid-derived monolayer to study bacteriophage translocation

Le,

Lubian,

Bowring

et al. 2024

Gut Microbes

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Bacteriophages (phages) are estimated to be the most abundant microorganisms on Earth. Their presence in human blood suggests that they can translocate from non-sterile sites such as the gastrointestinal tract where they are concentrated. To examine phage translocation ex vivo , we adapted a primary colonoid monolayer model possessing cell diversity and architecture, and a thick layer of mucus akin to the colonic environment in vivo . We show that the colonoid monolayer is superior to the Caco-2 cell-line model, possessing intact and organized tight junctions and generating a physiologically relevant mucus layer. We showed, using two different phages, that translocation across the colonoid monolayer was largely absent in differentiated monolayers that express mucus, unlike Caco-2 cultures that expressed little to no mucus. By stimulating mucus production or removing mucus, we further demonstrated the importance of colonic mucus in preventing phage translocation. Finally, we used etiological drivers of gut permeability (alcohol, fat, and inflammatory cytokines) to measure their effects on phage translocation, demonstrating that all three stimuli have the capacity to amplify phage translocation. These findings suggest that phage translocation does occur in vivo but may be largely dependent on colonic mucus, an important insight to consider in future phage applications.

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Section: Methodsmentioning

confidence: 99%