Page 74 - ebook

[B. Cell Biology/Stem Cell] B-37

Functional human small intestinal epithelium and their

applications in human microbiome research

Ohman Kwon¹ , KwangBo Jung¹,² , Mooseung Lee¹,², Dae-Soo Kim¹,², Mi-Young Son¹,²*
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¹Stem Cell Research Convergence Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB),

Daejeon 34141, Korea, ²KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon

34113, Korea

Advanced technologies are required for generating human intestinal epithelial cells (hIECs) harboring cellular
diversity and functionalities to predict drug absorption in humans and study normal intestinal epithelial physiology.

We developed a reproducible two-step protocol to induce human pluripotent stem cells to differentiate into a
functional hIEC monolayer exhibiting intestinal molecular features, cell type diversity, and high activities of intestinal

transporters and metabolic enzymes such as cytochrome P450 3A4 (CYP3A4). Functional hIECs are more suitable
for predicting compounds metabolized by CYP3A4 and absorbed in the intestine than Caco-2 cells. A significantly

higher number of bacteria were in contact with the apical surface of the functional hIEC monolayers. Moreover, a
higher number of macrophages attached to the basolateral side of the functional hIEC monolayer and translocated

to the upper side of the Transwell compared to that in immature hIECs, which may be attributed to elevated receptor
expression of bacterial antigens. This system is a step toward the transition from 3D intestinal organoids to 2D hIEC

monolayers without compromising cellular diversity and function. A physiologically relevant hIEC model offers a
novel platform for creating patient-specific assays and support translational applications, thereby bridging the gap
between 3D and 2D culture models of the intestine.