Co-culture model of Caco-2/HT29-MTX cells: A promising tool for investigation of phycotoxins toxicity on the intestinal barrier.

Most lipophilic phycotoxins have been involved in human intoxications but some of these toxins have never been proven to induce human gastro-intestinal symptoms, although intestinal damage in rodents has been documented. For investigating the in vitro toxicological profile of lipophilic phycotoxins on intestine, the epithelial Caco-2 cell line has been the most commonly used model. Nevertheless, considering the complexity of the intestinal epithelium, in vitro co-cultures integrating enterocyte-like and mucus-secreting cell types are expected to provide more relevant data. In this study, the toxic effects (viability, inflammation, cellular monolayer integrity, modulation of cell type proportion and production of mucus) of four lipophilic phycotoxins (PTX2, YTX, AZA1 and OA) were evaluated in Caco-2/HT29-MTX co-cultured cells. The four toxins induced a reduction of viability from 20% to 50% and affected the monolayer integrity. Our results showed that the HT29-MTX cells population were more sensitive to OA and PTX2 than Caco-2 cells. Among the four phycotoxins, OA induced inflammation (28-fold increase of IL-8 release) and also a slight increase of both mucus production (up-regulation of mucins mRNA expression) and mucus secretion (mucus area and density). For PTX2 we observed an increase of IL-8 release but weaker than OA. Intestinal cell models integrating several cell types can contribute to improve hazard characterization and to describe more accurately the modes of action of phycotoxins.