Co-cultures of enterocytes and hepatocytes for retinoid transport and metabolism

• Published in: Toxicology in vitro 2012-12, Vol.26 (8), p.1256-1264
• Main Authors: Rossi, Carlotta, Guantario, Barbara, Ferruzza, Simonetta, Guguen-Guillouzo, Christiane, Sambuy, Yula, Scarino, Maria Laura, Bellovino, Diana
• Format: Article
• Language: English
• Subjects: 3A cells; Animals; beta Carotene - pharmacokinetics; Biological Transport; Caco-2/TC7; Cell Differentiation; Cell Line; Coculture Techniques; CYP26A1; Cytochrome P-450 Enzyme System - genetics; Enterocytes - metabolism; HepaRG; Hepatocytes - metabolism; Humans; Mice; Models, Biological; Retinoic Acid 4-Hydroxylase; Retinol; Retinol Binding Protein 4; RNA Polymerase II - secretion; RNA, Messenger - metabolism; Up-Regulation - drug effects; Vitamin A - pharmacokinetics; β-carotene
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2012.04.013

► An in vitro co-culture system of human differentiated intestinal and hepatic cells is presented. ► The interplay between intestine and liver transport and metabolism was tested using dietary retinoids. ► Beta-carotene and retinol given to enterocytes acted on retinoid metabolism of co-cultured hepatocytes. ► For the homologous “all human” system we used highly differentiated human HepaRG hepatocytes. Dietary retinoid bioavailability involves the interplay of the intestine (transport and metabolism) and the liver (secondary metabolism). To reproduce these processes in vitro, differentiated human intestinal Caco-2/TC7 cells were co-cultured with two hepatocyte cell lines. Murine 3A cells and the more highly differentiated human HepaRG hepatocytes were both shown to respond to β-carotene (BC) and retinol (ROH) treatment by secreting Retinol Binding Protein 4 (RBP4). In co-culture experiments, Caco-2/TC7 were differentiated on filter inserts and transferred for the time of the experiment to culture wells containing confluent 3A or differentiated HepaRG cells. Functionality of the co-cultures was assayed using as endpoints the retinol-dependent secretion of RBP4 and the retinoic acid-dependent induction of CYP26A1 in hepatocytes. BC and ROH added to intestinal Caco-2/TC7 induced a reduction in intracellular RBP4 levels in the underlying hepatocytes and its secretion into the medium. HepaRG hepatocytes were also shown to up-regulate the expression of CYP26A1 mRNA in response to retinoid treatment. This in vitro model represents a useful tool to analyze the absorption and metabolism of retinoids and could be further developed to investigate other dietary compounds and molecules of pharmacological interest.