Miniaturisation and automatisation of Caco-2 permeability studies for screening of natural and synthetic ligands

Caco-2 cell monolayers have been widely accepted by pharmaceutical companies and by regulatory authorities as a standard in vitro–model system to predict permeability of compounds in human. To obtain these monolayers Caco-2 cells are traditionally grown on 12 and 24 wells for 21 to 28 days which is time consuming, laborious and expensive. To adapt the model to the needs of modern high-throughput screening, we replaced the 12- and 24-well plates with 96-well plates and reduced the growing time to 7 days. A set of standard compounds with different permeabilities, various permeability markers and confocal microscopy were used to assess the utility of our new method on Biomek FX automation. The permeability results obtained from standard compounds used were comparable to those obtained from traditionally performed experiments. These results indicate that we managed to build up a fast miniaturized and automated protocol to make a first evaluation of permeation of new compounds in the drug discovery process.

Acknowledgements: The European Commission 6th framework program Pro-Kinase Research project no. 503467