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Assessment of iron availability using combined in vitro digestion and Caco-2 cell culture


, : Assessment of iron availability using combined in vitro digestion and Caco-2 cell culture. Nutrition Research 16(3): 479-487

A model for the rapid assessment of iron availability was developed that combines in vitro digestion with iron uptake by Caco-2 cell monolayers. In this method, samples (beef, ascorbic acid, or citric acid) were adjusted to pH 2, labeled with 59Fe, and subjected to pepsin digestion (pH 2, 37 degree C) for 1 h to simulate gastric digestion. Next, a dialysis bag (12,000-14,000 molecular weight cutoff) containing 150 mM PIPES buffer (pH 6.7) was placed in the digest and incubation continued for 30 min. Then, a pancreatin-bile mixture was added, and incubation was continued for an additional 2 h. The contents of the dialysis bag were removed and an aliquot applied to Caco-2 cell monolayers. After a 60 min incubation, iron that was non-specifically bound to the surface of the monolayer was removed by rinsing with a solution containing bathophenanthroline disulfonic acid and sodium dithionite. Cells were then counted for 59Fe activity to measure uptake. Beef and ascorbic acid enhanced Caco-2 cell iron uptake, whereas citric acid had no effect. These results compare favorably with literature reports of human studies and suggest that a dialyzable factor(s) less than 14,000 daltons, released during beef digestion, was responsible for the iron absorption-enhancing properties of beef We believe that this system will be useful for studying basic mechanisms of iron absorption and for in vitro estimation of iron bioavailability.

US$19.90

DOI: 10.1016/0271-5317(96)00029-2


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