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Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers


Permeability coefficients across monolayers of the human colon carcinoma cell line Caco-2, cultured on permeable supports, are commonly used to predict the absorption of orally administered drugs and other xenobiotics. This protocol describes our method for the cultivation, characterization and determination of permeability coefficients of xenobiotics (which are, typically, drug-like compounds) in the Caco-2 model. A few modifications that have been introduced over the years are incorporated in the protocol. The method can be used to trace the permeability of a test compound in two directions, from the apical to the basolateral side or vice versa, and both passive and active transport processes can be studied. The permeability assay can be completed within one working day, provided that the Caco-2 monolayers have been cultured and differentiated on the permeable supports 3 weeks in advance.

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Figure 1
Figure 2: Application of the technique in absorption and drug metabolism studies.
Figure 3: Morphology of Caco-2 cell monolayers.
Figure 4: Anticipated results.


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This work has been supported by the Swedish Research Council, the Swedish Animal Welfare Agency and EU-grant no. LSHB-CT-2004-512051.

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Correspondence to Per Artursson.

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Hubatsch, I., Ragnarsson, E. & Artursson, P. Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers. Nat Protoc 2, 2111–2119 (2007).

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