US EPA

214604 

Journal Article 

A modified fast (4 day) 96-well plate Caco-2 permeability assay 

Uchida, M; Fukazawa, T; Yamazaki, Y; Hashimoto, H; Miyamoto, Y 

2009 

Yes 

Journal of Pharmacological and Toxicological Methods
ISSN: 1056-8719 

59 

1 

39-43 

English 

19049886 

10.1016/j.vascn.2008.10.006 

INTRODUCTION: The Caco-2 permeability assay is widely used for lead optimization in drug discovery. A 3 to 5-day system using a 24-well plate and a 10 to 21-day system using a 96-well plate have been established. Here, we modified the assay system to provide a ready-to-use Caco-2 cell monolayer using a 96-well plate in just 4 days. METHODS AND RESULTS: In our system, collagen-coated inserts and the prolongation of the culture period after seeding leads to greater Caco-2 cell proliferation and sufficient contact-inhibition. The differentiation of Caco-2 cells was enhanced, when the contact-inhibited Caco-2 cells were exposed to the differentiation-inducing agent butyric acid. The permeability to nine well-known compounds showed a statistical correlation between our 4-day system using a 96-well plate and the conventional 21-day system using a 24-well plate. DISCUSSION: We conclude that our system is more useful for evaluating many compounds for lead optimization in drug discovery. 

Butyric Acid/pd [Pharmacology]; Caco-2 Cells; Cell Culture Techniques; Cell Differentiation/de [Drug Effects]; Cell Membrane Permeability/de [Drug Effects]; Cell Proliferation/de [Drug Effects]; Cells, Cultured; Coated Materials, Biocompatible; Humans; Pharmaceutical Preparations/ch [Chemistry]; Pharmaceutical Preparations/cl [Classification]; Pharmaceutical Preparations/me [Metabolism]; Propranolol/pd [Pharmacology]; Time Factors; Vinblastine/pd [Pharmacology]; 0 (Coated Materials, Biocompatible); 0 (Pharmaceutical Preparations); 107-92-6 (Butyric Acid); 525-66-6 (Propranolol); 865-21-4 (Vinblastine)