Standardization of Caco-2 cell culture as in vitro model for intestinal permeability

Abstract

The aim of this study was to find out the optimal experimental conditions for Caco-2 cell culture (time and density) and permeability assays (diffusion system and drug concentration) in order to study the in vitro drugs permeability as a predictive method for drug absorption across intestinal epithelium. The integrity of the monolayers used in each assay was determined by measuring the transepithelial electrical resistance (TEER) and the permeability of the atenolol-a drug which is transported across the monolayers by the paracellular pathway-. The best working condition was obtained with a cell seeding of 7.10⁴ cells/insert in a vertical difussion chamber. In such context, the monolayers had a TEER higher than 550 Ω.cm² and the apparent permeability coefficient of atenolol was 0.71 ± 0.19 x 10^-6 cm/seg.