Proposing a Caco-2/HepG2 cell model for in vitro iron absorption studies.

J Nutr Biochem

Chalmers University of Technology, Department of Chemical and Biological Engineering, Life Science division, Food Science, SE-412 96 Gothenburg, Sweden.

Published: July 2014

AI Article Synopsis

  • The study enhances the Caco-2 cell line model for iron absorption by including human liver cells (HepG2), improving its ability to replicate the regulation by hepcidin.
  • In co-cultured Caco-2 cells with HepG2, ferritin levels and iron transport across the layers increased significantly compared to monocultured Caco-2 cells.
  • The research also found that sourdough bread boosts iron bioavailability more effectively than heat-treated bread, highlighting the importance of considering hepatic regulation in iron absorption studies.

Article Abstract

The Caco-2 cell line is well established as an in vitro model for iron absorption. However, the model does not reflect the regulation of iron absorption by hepcidin produced in the liver. We aimed to develop the Caco-2 model by introducing human liver cells (HepG2) to Caco-2 cells. The Caco-2 and HepG2 epithelia were separated by a liquid compartment, which allowed for epithelial interaction. Ferritin levels in cocultured Caco-2 controls were 21.7±10.3 ng/mg protein compared to 7.7±5.8 ng/mg protein in monocultured Caco-2 cells. The iron transport across Caco-2 layers was increased when liver cells were present (8.1%±1.5% compared to 3.5%±2.5% at 120 μM Fe). Caco-2 cells were exposed to 0, 80 and 120 μM Fe and responded with increased hepcidin production at 120 μM Fe (3.6±0.3 ng/ml compared to 2.7±0.3 ng/ml). The expression of iron exporter ferroportin in Caco-2 cells was decreased at the hepcidin concentration of 3.6 ng/ml and undetectable at external addition of hepcidin (10 ng/ml). The apical transporter DMT1 was also undetectable at 10 ng/ml but was unchanged at the lower concentrations. In addition, we observed that sourdough bread, in comparison to heat-treated bread, increased the bioavailability of iron despite similar iron content (53% increase in ferritin formation, 97% increase in hepcidin release). This effect was not observed in monocultured Caco-2 cells. The Caco-2/HepG2 model provides an alternative approach to in vitro iron absorption studies in which the hepatic regulation of iron transport must be considered.

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http://dx.doi.org/10.1016/j.jnutbio.2014.02.013DOI Listing

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