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.
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.
Authors: David Bruhn; Dora B Madhura; Marcus Maddox; Robin B Lee; Ashit Trivedi; Lei Yang; Michael S Scherman; Janet C Gilliland; Veronica Gruppo; Michael R McNeil; Anne J Lenaerts; Bernd Meibohm; Richard E Lee Journal: Bioorg Med Chem Date: 2012-08-28 Impact factor: 3.641
Authors: Hye-Sik Kong; Shuo Tian; Yali Kong; Guanhua Du; Li Zhang; Mira Jung; Anatoly Dritschilo; Milton L Brown Journal: Pharm Res Date: 2012-07-27 Impact factor: 4.200