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Experimental Read Out

Functionality Studies

HT29 cells have been widely employed to assess the potential anticancer effect of different food compounds, and their gastrointestinal digests, on the basis of the carcinoma origin of this cell line. An illustrative example is found in the study of the antiproliferative activity of the in vitro gastrointestinal digest of sea cucumber wall (Pérez-Vega et al. 2013). Another representative study is found in the assessment of the cytotoxic activity of different extracts from “Racimo” tomato variety and simulated digests against HT29 cell growth, including also the evaluation of the selectivity of the toxic effect against cancer cells (Guil-Guerrero et al. 2011). Other activities have been widely studied on HT29 cells or derived lines, such as immunomodulatory, antioxidant or barrier protective properties. For instance, milk and soy ferments with different strains of lactic acid bacteria (LAB) were evaluated on human intestinal epithelial cells, including HT29 cells, to test their immunomodulatory activity (Wagar et al. 2009). The study was carried out on cells treated with TNF-α and the production of IL-8 was evaluated in the cell supernatant by a commercial enzyme-linked immunosorbent assay. Another investigation focused on the study of the immunomodulatory activity of cereal β-glucan preparations on both HT29 and Caco-2 cells (Rieder et al. 2011). In vitro data of antioxidative activity for food compounds is often presented together with genotoxic tests performed with HT29 cells. Among others, the comet assay has become one of the standard methods for assessing DNA integrity (Collins 2004). Ferguson et al. (2005) demonstrated both in vitro antioxidant activity and antigenotoxic effect in HT29 cells by using a comet assay for two hydroxycinnamic acids with an important percentage in certain plant foods, as spinach or cereals. Interestingly, the protection of resveratrol and quercetin against exogenous pro-oxidative damage was determined in HT29 cells with induced oxidative stress by addition of fatty acid hydroperoxides (Kaindl et al. 2008). HT29-derived cell lines constitute a valuable tool related to the strengthening of the intestinal mucus barrier to study the mucin-stimulating activity of food compounds. The protocol described above has been recently applied to asses the mucinstimulating effect of certain milk protein hydrolysates and derived peptides (Martínez-Maqueda et al. 2012, 2013a, b). HT29-MTX cells and an analogous experimental design was also implemented to evaluate the regulation of the mucin production by the β-casomorphin-7, a μ-opioid peptide derived from bovine milk (Zoghbi et al. 2006), or by a yoghurt peptide pool (Plaisancié et al. 2013). Other HT29 derived cell lines have been employed due to their capability to form a mucuslayer, e.g. HT29/B6 cells differentiated in a glucose-free culture (Kreusel et al. 1991). Hering et al. (2011) evaluated the expression of tight junction proteins and the related intestinal barrier-protection in HT29/B6 cells, under the effect of TGF-β, a whey protein component. Likewise, monoterpene d-limonene, naturally occurring in the rind of citrus fruit, showed a significant increase on the transepithelial resistance in HT29/B6 cells (D'Alessio et al. 2013).

 
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