Automated absorption assessment using Caco-2 cells cultured on both sides of polycarbonate membranes

Purpose. To increase the capacity of in vitro absorption assessment and to decrease the amount of substance needed to perform early mechanistic invest igations. Methods. A liquid handling system, combined with a shaker and heating plate s. was used to automate the Caco-2 cell based in vitro absorption assessmen t assay. In order to decrease the amount of substance needed for early mech anistic studies, a method for culturing Caco-2 cells on the lower side of p olycarbonate membranes was also developed. Results. Similar results were obtained with the automated assay as compared to manually performed assays. Data presented suggest that active transport and efflux were decreased in cells cultured on the lower side of the membr anes as compared to ordinary seeded cells. Conclusions. Implementation of a liquid handling system for in vitro absorp tion assessment as reported here decrease the manual workload and increases the capacity of this in vitro assay substantially. Caco-2 cells cultured o n the lower side of polycarbonate membranes, as described in this article, can not be used for analysis of transport mechanisms.