Monolayer platform using human biopsy-derived duodenal organoids for pharmaceutical research

Mol Ther Methods Clin Dev. 2021 May 19:22:263-278. doi: 10.1016/j.omtm.2021.05.005. eCollection 2021 Sep 10.


The human small intestine is the key organ for absorption, metabolism, and excretion of orally administered drugs. To preclinically predict these reactions in drug discovery research, a cell model that can precisely recapitulate the in vivo human intestinal monolayer is desired. In this study, we developed a monolayer platform using human biopsy-derived duodenal organoids for application to pharmacokinetic studies. The human duodenal organoid-derived monolayer was prepared by a simple method in 3-8 days. It consisted of polarized absorptive cells and had tight junctions. It showed much higher cytochrome P450 (CYP)3A4 and carboxylesterase (CES)2 activities than did the existing models (Caco-2 cells). It also showed efflux activity of P-glycoprotein (P-gp) and inducibility of CYP3A4. Finally, its gene expression profile was closer to the adult human duodenum, compared to the profile of Caco-2 cells. Based on these findings, this monolayer assay system using biopsy-derived human intestinal organoids is likely to be widely adopted.

Keywords: CES1; CES2; CYP3A4; P-glycoprotein; PEPT1; intestinal first-pass effect; intestinal organoids; microarray; monolayer culture; small intestine.