Micronutrient deficiencies are prevalent worldwide and have detrimental effects on human health. Complex interactions between micronutrients and other dietary components largely determine micronutrient bioavailability, and understanding these interactions is key to improving micronutrient status. A number of in vitro and in vivo methodologies are available for assessing micronutrient bioavailability. The purpose of this review is to highlight the usefulness of one of the in vitro models, the Caco-2 cell, as a predictive tool for human micronutrient bioavailability. The review focuses on current methods used with the Caco-2 cell line, their benefits and limitations, and the possibilities for the future development of this model.