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Updated: Jun 27, 2026
Using Caco-2 Cells to Study Lipid Transport by the Intestine
Published on: August 20, 2015
A modified fast (4 day) 96-well plate Caco-2 permeability assay
Masashi Uchida1, Tominaga Fukazawa, Yuri Yamazaki
1Toxicology and Pharmacokinetics Laboratories, Pharmaceutical Research Laboratories, Toray Industries, Inc, Kamakura, Kanagawa, Japan. Masashi_Uchida@nts.toray.co.jp
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.
