T84 monolayers are superior to Caco-2 as a model system of colonocytes

Abstract

Colonic adenocarcinoma-derived Caco-2 and T84 epithelial cell lines are frequently used as in vitro model systems of functional epithelial barriers. Both are utilised interchangeably despite evidence that differentiated Caco-2 cells are more reminiscent of small intestinal enterocytes than of colonocytes, whereas differentiated T84 cells are less well characterised. The aim of this study was, therefore, to further characterise and compare differentiated Caco-2 and T84 cells. The objectives were to (1) compare the brush border morphology, (2) measure the expression of enterocyte- and colonocyte-specific genes and (3) compare their response to butyrate, which is dependent on the monocarboxylate transporter 1 (MCT1), an apical protein expressed primarily in colonocytes. T84 microvilli were significantly shorter than those of Caco-2 cells, which is a characteristic difference between small intestinal enterocytes and colonocytes. Also, enterocyte-associated brush border enzymes expressed in differentiated Caco-2 cells were not increased during T84 maturation, whereas colonic markers such as MCT1 were more abundant in differentiated T84 cells compared to differentiated Caco-2 cells. Consequently, T84 cells displayed a dose-responsive improvement of barrier function towards butyrate, which was absent in Caco-2 cells. On the other hand, differences in epithelial toll-like receptor expression between Caco-2 and T84 monolayers did not result in a corresponding differential functional response. We conclude that differentiated Caco-2 and T84 cells have distinct morphological, biochemical and functional characteristics, suggesting that T84 cells do not acquire the biochemical signature of mature small intestinal enterocytes like Caco-2 cells, but retain much of their original colonic characteristics throughout differentiation. These findings can help investigators select the appropriate intestinal epithelial cell line for specific in vitro research purposes.

Serotonin is an endogenous regulator of intestinal CYP1A1 via AhR

“…Caco-2 cells are derived from the large intestine, but functionally and morphologically represent enterocytes upon differentiation. On the other hand, the colonic crypt-derived cell line T84 has characteristics similar to colonocytes 35 . To determine if the effect of 5-HT on CYP1A1 mRNA was cell-line specific, T84 cells were treated with varying concentrations of 5-HT (10–100 μM 5-HT).…”

Gut Microbiome, Intestinal Permeability, and Tissue Bacteria in Metabolic Disease: Perpetrators or Bystanders?

“…This impairment was further exacerbated by lipid challenge in samples emanating from subjects with obesity as compared with lean subjects and was an independent explanatory variable for the presence of T2D [92]. Changes to the tight junction proteins from the claudin and zonula occludens family have also been observed with reduction in occludin and tricellulin [93] in obesity. This has been similarly shown in mice, where HFD induced a shift in claudin expression [94], as well as in hyperleptinemic db/db mice, which displayed a decreased intestinal resistance and a profound modification of the occludin and ZO-1 expression in their intestinal mucosa [95].…”

“…This has been similarly shown in mice, where HFD induced a shift in claudin expression [94], as well as in hyperleptinemic db/db mice, which displayed a decreased intestinal resistance and a profound modification of the occludin and ZO-1 expression in their intestinal mucosa [95]. This observation was accompanied by higher circulating levels of tumor necrosis α (TNF-α) and interferon γ (INF-γ), which have independently been shown to increase occludins and the internalization of claudin-1 and -4 and to downregulate expression of claudin-1, causing increased permeability in colonic adenocarcinoma-derived T84 epithelial cell lines [93]. In the link between obesity and increased permeability, leptin could play a pivotal role.…”

Enteropathogenic Escherichia coli (EPEC) Recruitment of PAR Polarity Protein Atypical PKCζ to Pedestals and Cell–Cell Contacts Precedes Disruption of Tight Junctions in Intestinal Epithelial Cells

“…Although all derived from colonic cancer cell lines, Caco-2 when grown for extended periods differentiate into small intestine-like cells, whereas SKCO-15 and T84 cells are more colonic in nature. Several studies have focused on the biochemical and structural differences between these colonic cell lines [58][59][60]. In addition, SNX9 expression levels differ between colon cancer cell lines, as well as having other varying redundant sorting nexin proteins [61][62][63].…”