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BMC Cell Biol. 2006 May 2;7:20.

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

Author information

1
diGESD (Study Group of Digestive System), Center of Immunology, Institute of Pathology, University of Liege, 4000 Liege, Belgium. g.nollevaux@kitozyme.com

Abstract

BACKGROUND:

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.

RESULTS:

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.

CONCLUSION:

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.

PMID:
16670004
PMCID:
PMC1617214
DOI:
10.1186/1471-2121-7-20
[Indexed for MEDLINE]
Free PMC Article
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