JoVE Encyclopedia of Experiments
Biological Techniques
0 views • 3:56 min • July 8th, 2025
Enterocytes are specialized small intestine cells that play a crucial role in nutrient uptake, including iron absorption.
To assess iron bioavailability - the proportion of ingested iron accessible for physiological functions - take an acidified solution containing the food sample. Now, add pepsin - a protein digestion enzyme, from the stomach. At acidic pH, pepsin initiates gastric digestion by breaking down complex dietary proteins into small peptides.
Add sodium bicarbonate to increase the pH and stop gastric digestion. Add a solution containing pancreatic enzymes and bile to simulate intestinal digestion. Transfer the mixture into the upper portion of a two-chamber cell culture system.
The lower chamber contains enterocytes differentiated from Caco-2 - a colon adenocarcinoma cell line. A dialysis membrane separates the upper chamber from the lower one. The membrane allows the diffusion of low molecular weight species.
The pancreatic enzymes-bile mixture in the upper chamber breaks down carbohydrates, proteins, and lipids into smaller subunits - releasing iron in the form of ferric ions.
Bioavailability enhancers in the food convert the released ferric ions to the ferrous state
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