Use of a combinedin vitroassay for effect-directed assessment of infant formulas

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Milk (M), soya (S) and hypoallergenic (HA) infant formulas were evaluated for an effect-directed assessment of whole food on human liver (HuH6) and colon (DLD-1) cell lines. Treatment concentrations were derived from an estimated meal for a 4-month infant. The expression of a panel of genes (ERα, ERβ, AR, PPARγ, AhR) and functional markers (TNFα, IL-8 in DLD1; γ-GT in HuH6) were evaluated comparing results with bisphenol A, vinclozolin, genistein, TCDD and mixture effects at concentrations reflecting dietary exposures. M increased only ERα in DLD-1; HA affected only DLD-1 (ERα and ERβ upregulation; AhR and TNFα downregulation); S upregulated ERβ both cell lines), ERα (DLD-1) and AR (HuH6). Bisphenol A showed similar effects as HA and S in DLD-1; the mixture upregulated AR and ERβ in HuH6 as S. Although DLD-1 was more responsive than HuH6, the use of both cell lines and all selected endpoints allow to characterise each tested formula or substance.


The in vitro test battery, consisting of two human cell lines from liver (HuH6) and colon (DLD-1), allows to assess potential EDs-like effects of three commercial infant formulas (as shown for milk formula) through biomarkers evaluation (i.e. nuclear receptor gene expression and protein modulation). The results point out that each infant formula alters a distinct pattern of biomarkers in relation to each human cell line used.

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