When looking at Caco-2 cell cultures microscopically, it is evident even by visual inspection that the cells are heterogeneous. As a result, over the years the characteristics of the cells used in different laboratories around the world have diverged significantly, which makes it difficult to compare results across labs.[4]
Caco-2 cells are most commonly used not as individual cells, but as a confluent monolayer on a cell culture insert filter (e.g., Transwell). When cultured in this format, the cells differentiate to form a polarized epithelial cell monolayer that provides a physical and biochemical barrier to the passage of ions and small molecules.[3][5] The Caco-2 monolayer is widely used across the pharmaceutical industry as an in vitro model of the human small intestinal mucosa to predict the absorption of orally administered drugs. The correlation between the in vitro apparent permeability (P¬app) across Caco-2 monolayers and the in vivo fraction absorbed (fa) is well established