Intestinal Epithelium Model

As long-term primary culture of human intestinal epithelial cells is still a challenge, most intestinal epithelial systems rely on immortalized cell lines (Grossmann et al., 1998). In order to clarify the ability of yeast and filamentous forms to influence adherence of C. albicans to the intestinal epithelium, the enterocyte cell lines Caco-2 and HT29 were tested. Candida-enterocytes interaction was observed with high-resolution scanning electron microscopy, while yeast adherence to enterocytes was quantified by using an ELISA assay (Wiesner et al., 2002) concluding that both C. albicans morphologies, yeast, and hyphae, could adhere to (and perhaps invade)

the apical surface of cultured enterocytes. Caco-2 cells have been also used to demonstrate that p, 1-2 and a, 1-2 oligomannosides mediate the adherence of C. albicans blastospores to human enterocytes (Dalle et al., 2003). Adherence was quantified by immunofluorescence and the percentage of adhesion was determined as the ratio of the number of adherent yeasts on the entire surface of the coverslip to the inoculum.

The Caco-2 line proved to give a good performance in a reconstituted intestinal epithelium model in comparison to HT29 and Lovo cells as determined by Dieterich et al. (2002). Caco-2 cells display a regularly ordered single cell layer which differentiates microvilli on the apical membrane. To construct the three-dimensional intestinal model cells were grown on a collagen matrix supplemented with primary fibroblasts and myoblasts. This model has been useful to define the defects on adhesion and invasion of the avirulent C. albicans cph1 and efg1 mutant, showing that double efg1 cph1 mutants were unable to adhere or penetrate neither of the model systems tested (Dieterich et al., 2002).

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