Establishment of human intestinal organoids derived from commercially available cryopreserved intestinal epithelium and evaluation for pharmacokinetic study

Abstract

Human intestinal organoids (HIOs) have been reported to exert their functions in a way that mimics living organs, and HIOs-derived monolayers are expected to be applied to in vitro intestinal pharmacokinetic studies. However, HIOs are established from human tissue, which raises issues of availability and ethics. In the present study, to solve these problems, we have established intestinal organoids using commercially available cryopreserved human intestinal epithelial cells (C–IOs), and compared their functions with biopsy-derived human intestinal organoids (B–IOs) from a pharmacokinetic point of view. Both C–IOs and B–IOs reproduced the morphological features of the intestinal tract and were shown to be composed of epithelial cells. Monolayers generated from C–IOs and B–IOs (C-IO-2D, B-IO-2D, respectively) structurally mimic the small intestine. The C–IOs showed gene expression levels comparable to those of the B–IOs, which were close to those of adult human small intestine. Importantly, the C–IOs-2D showed levels of pharmacokinetics-related protein expression and activity—including cytochrome P450 3A4 (CYP3A4) and carboxylesterase 2 (CES2) enzymatic activities and P-glycoprotein (P-gp) transporter activities —similar to those of B–IOs-2D. This study addresses the difficulties associated with B–IOs and provides fundamental characteristics for the application of C–IOs in pharmacokinetic studies.