Development of the method for evaluating protective effect of food factors on THP‐1‐induced damage to human intestinal Caco‐2 monolayers

Abstract

Immune cells located in the intestinal epithelium interact with intestinal epithelial cells via soluble factors. In this study, a new in vitro model using a coculture system was constructed to analyze the interaction between intestinal epithelial cells and macrophage-like cells. Human intestinal epithelial Caco-2 cells were differentiated on semipermeable membranes. Human monocytic THP-1 cells were differentiated to macrophage-like cells and then cocultured on the basolateral side of the Caco-2 cell monolayers. By coculturing for 48 hours, an increased release of lactate dehydrogenase from the Caco-2 cells and a decrease in the transepithelial electrical resistance of the monolayers were observed, suggesting that the coculture with THP-1 induced some disruption of the Caco-2 cells. This disruption was significantly suppressed by adding the anti-TNF-alpha antibody to the medium, suggesting that TNF-alpha secreted from THP-1 caused damage to the Caco-2 cells. It is also suggested that this phenomenon is similar to that observed with inflammatory bowel disease (IBD). The effects of food factors on the cells in this coculture system were examined. The disruption of the Caco-2 cell monolayers was significantly reduced by adding caffeine to the medium on the apical side. It is hoped that this coculture system will be a good model for the treatment of IBD.

Induction by activated macrophage-like THP-1 cells of apoptotic and necrotic cell death in intestinal epithelial Caco-2 monolayers via tumor necrosis factor-alpha

“…We have previously reported the interaction between Caco-2 cells as a model of human intestinal epithelial cells and PC12 cells as a model of neuronal cells [24,25]. We have also reported preliminary results on the interaction between intestinal epithelial cells and immune cells [26].…”

Cytoplasmic Form of Carlr lncRNA Facilitates Inflammatory Gene Expression upon NF-κB Activation

“…Both of these genes are important players in inflammatory intestinal diseases and their activation induces secretion of prostaglandin and the IL1β inflammatory cytokine by THP-1 cells, soluble factors that modulate interactions between epithelial and immune cells. To further analyze the relationship between THP-1 and intestinal cells, we utilized a co-culture model of intestinal inflammation (27). After co-culturing LPS-stimulated THP-1 cells with intestinal Caco2 cells, we observed an increase in proliferation of the intestinal cells (Figure 4B) that was diminished if Carlr was previously silenced in THP-1 cells (Figure 4C).…”

Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal

“…Although not mimicking the ageing gut epithelium, FHs 74 Int and Caco-2 cell lines are employed world wide as suitable cell culture models displaying enterocyte like differentiation for studying the proliferation and gut integrity/permeability and immunomodulation at cellular level [ 23 - 30 , 32 , 33 ]. We first determined a concentration range of Enprocal, which was not cytotoxic (kill/disrupt) to the Caco-2 cell monolayers, needed to test the downstream intestinal permeability assays, adhesion assays etc.…”

“…Recently an in vitro coculture system has been reported that could be utilized for an assay to search for drugs or food substances, which could prevent intestinal inflammation [ 33 ]. To study the cell-to-cell interaction between intestinal epithelial cells (Caco-2) and activated macrophages (THP-1), we utilized the similar Transwell model.…”

“…We employed a coculture model to examine the effect of Enprocal D treatment on the interaction between gut-immune cells. For the coculture experiments THP-1 monocytic cells were grown in 24-well plates at a density of 5 × 10 5 cells/well and differentiated into macrophage-like cells, by treatment with 200 nM phorbol myristrate acetate (PMA; Sigma-Aldrich) for 4 days [ 33 ]. The differentiated status was confirmed on the basis of morphology and staining for the expression of leukocyte integrin CD11b (macrophage-1 antigen) (data not shown).…”