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

Autor: Schneider Yves-Jacques, Deloyer Patricia, Zorzi Willy, El Moualij Benaïssa, Devillé Christelle, Nollevaux Géraldine, Peulen Olivier, Dandrifosse Guy
Jazyk: angličtina
Rok vydání: 2006
Předmět:
Zdroj: BMC Cell Biology, Vol 7, Iss 1, p 20 (2006)
Druh dokumentu: article
ISSN: 1471-2121
DOI: 10.1186/1471-2121-7-20
Popis: 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.
Databáze: Directory of Open Access Journals