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Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21)

Géraldine Nollevaux1, Christelle Devillé1, Benaïssa El Moualij2, Willy Zorzi2, Patricia Deloyer1, Yves-Jacques Schneider3, Olivier Peulen1 and Guy Dandrifosse1*

Author affiliations

1 diGESD (Study Group of Digestive System), Center of Immunology, Institute of Pathology, University of Liege, 4000 Liege, Belgium

2 Department of human histology, Center of Immunology, CRPP, Institute of Pathology, University of Liege CHU, 4000 Liege, Belgium

3 Department of Cellular Biochemistry, Catholic University of Louvain, 1348 Louvain-la-Neuve, Belgium

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Citation and License

BMC Cell Biology 2006, 7:20  doi:10.1186/1471-2121-7-20

Published: 2 May 2006



The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure.


In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used (serum-free medium) did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells.


The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules.