Abstract
Drug absorption via the intestinal tissue is modulated by membrane permeability and metabolism in intestinal epithelial cells (IECs). In drug discovery research, using human IECs to evaluate membrane permeability and metabolic stability can offer very useful information when exploring for drug candidate compounds that have good bioavailability and when trying to predict the fraction absorbed and intestinal availability in humans. Here, we evaluated the pharmacokinetic functions of human IECs differentiated from human induced pluripotent stem cells (hiPSCs) in 3D cultures. As human IECs differentiated in 3D cultures form intestinal organoids and spheroids (herein termed organoids), their morphology makes it difficult to evaluate their pharmacokinetic functions. Therefore, we dissociated intestinal organoids into single cells and attempted to purify human IECs. We found that hiPSC-derived IECs (hiPSC-IECs) expressed the epithelial cell adhesion molecule (EpCAM) and could be highly purified by sorting EpCAM+ cells. The hiPSC-IEC monolayer showed a high TEER value (approximately 350 Ω × cm2). In addition, hiPSC-IECs oxidatively metabolized terfenadine (CYP3A and CYP2J2 substrate) and midazolam (CYP3A substrate). These results indicated that hiPSC-IECs form tight-junction and have cytochrome P450 enzymatic activities. In conclusion, we developed a novel application of hiPSC-derived intestinal organoids for drug testing.
Highlights
Drug absorption via the intestinal tissue is modulated by membrane permeability and metabolism in intestinal epithelial cells (IECs)
To confirm the human induced pluripotent stem cells (hiPSCs)-IECs sorted by magnetic-activated cell sorting system (MACS), we examined the ratios of epithelial cell adhesion molecule (EpCAM)+ cells in the pre-separation cells, the positive and negative fractions after MACS by flow cytometric analysis
The value in hiPSC-IECs was significantly higher compared to those of the pre-separation cells and the EpCAM negative cells (p < 0.01). These results revealed that the purified hiPSC-IECs exhibited CYP3A and CYP2J2 metabolizing enzyme activities, with a high activity for CYP2J2 that is responsible for the hydroxylation reaction of terfenadine (Fig. 3B)
Summary
Drug absorption via the intestinal tissue is modulated by membrane permeability and metabolism in intestinal epithelial cells (IECs). As in vitro high-throughput membrane permeation screening tools for drug candidate compounds, the parallel artificial membrane permeation assay and Caco-2 cell lines derived from human colorectal adenocarcinoma have been used[1,2,3,4] These evaluation tools are used to predict the fraction absorbed in humans. We aimed at developing a novel application of hiPSC-derived intestinal organoids for evaluation of membrane www.nature.com/scientificreports permeation and metabolism of drug candidate compounds. For this purpose, we attempted to dissociate intestinal organoids into single cells and purified hiPSC-IECs using an epithelial cell surface marker EpCAM11,12
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
