Protocol abstract
Nature Protocols 2, - 2111 - 2119 (2007)
Published online: 23 August 2007 | doi:10.1038/nprot.2007.303
Subject Categories: Cell and tissue culture | Utilizing model organisms
Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers
Ina Hubatsch1, Eva G E Ragnarsson1 & Per Artursson1
Abstract
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.
- Department of Pharmacy, Uppsala University, Box 580, SE-751 23 Uppsala, Sweden.
Correspondence to: Per Artursson1 e-mail: per.artursson@farmaci.uu.se
nature-products
A-Z product listing
- 12-well cell culture clusters with lids(Corning Costar Transwell)
- 12-well plates(Millipore)
- Bovine serum albumin(Sigma)
- Caco-2 cells(ATCC)
- Evohm epithelial voltmeter(WPI)
- Filters(Corning Costar Transwell)