Generation of Intestinal Organoids Suitable for Pharmacokinetic Studies from Human Induced Pluripotent Stem Cells

Abstract

Intestinal organoids morphologically resemble intestinal tissues and are expected to be used in both regenerative medicine and drug development studies, including pharmacokinetic studies. However, the pharmacokinetic properties of these organoids remain poorly characterized. In this study, we aimed to generate pharmacokinetically functional intestinal organoids from human induced pluripotent stem (iPS) cells. Human iPS cells were induced to differentiate into the midgut and then seeded on EZSPHERE plates (AGC Techno Glass Inc., Shizuoka, Japan) to generate uniform spheroids, and the floating spheroids were subsequently differentiated into intestinal organoids using small-molecule compounds. Exposure to the small-molecule compounds potently increased the expression of intestinal markers and pharmacokinetic-related genes in the organoids, and the organoids also included various intestinal cells such as enterocytes, intestinal stem cells, goblet cells, enteroendocrine cells, Paneth cells, smooth muscle cells, and fibroblasts. Moreover, microvilli and tight junctions were observed in the organoids. Furthermore, we detected not only the expression of drug transporters but also efflux transport activity through ABCB1/MDR1 and the induction of the drug-metabolizing enzyme CYP3A4 by ligands of nuclear receptors. Our results demonstrated the successful generation of pharmacokinetically functional intestinal organoids from human iPS cells. Thus, these intestinal organoids could be used as a pharmacokinetic evaluation system in drug development studies.

Organoid technology in disease modelling, drug development, personalized treatment and regeneration medicine

“…The technology to establish intestinal organoids have been developed [ 9 , 43 – 47 ]. IPSCs-derived intestinal organoids contained diverse intestinal cells, including intestinal stem cells, enterocytes, goblet cells, enteroendocrine cells, paneth cells, fibroblasts and smooth muscle cells [ 47 ]. In addition, intestinal markers and pharmacokinetic-related genes were detected, and some structural features (microvilli and tight junctions) were observed in organoids [ 47 ].…”

Establishment of a novel culture method for maintaining intestinal stem cells derived from human induced pluripotent stem cells

“…It was reported that use of three-dimensional (3D) cultures extended the period during which intestinal cells can be cultured (Jung et al, 2011;Sato et al, 2011Sato et al, , 2009. Moreover, the organoids in 3D cultures display a villus-like structure similar to intestinal tissue and contain several cells that are consistent with the crypt niche of the intestines (Sawant-Basak et al, 2018;Onozato et al, 2018;Tamminen et al, 2015;Foulke-Abel et al, 2014;Jung et al, 2011;Spence et al, 2011;Sato et al, 2011Sato et al, , 2009. Although stem cell characteristics can reportedly be maintained by mimicking the environment and structure of the living intestine, the exchange and passage of medium in 3D cultures are complicated.…”

Emerging Models of Drug Metabolism, Transporters, and Toxicity

“…Li et al (2018a) reported processing and optimization of mucosal epithelium and cryopreserved enterocytes (Ho et al, 2017) to study intestinal metabolism, P450 induction, and gastrointestinal toxicity. Similarly, Iwao and colleagues have reported differentiating human iPSCs into functional enterocytes (Onozato et al, 2018). In this issue, an intestinal mini-review outlines applications and current limitations of such emerging human derived models bioengineered from intestinal crypts, mucosal extracts, iPSCderived organoids, and human intestinal tissue (Sawant-Basak et al, 2018).…”