Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

Abstract

Adult intestinal stem cells (ISCs) possess both a long-term proliferation ability and differentiation capability into enterocytes. As a novel in vitro system for the evaluation of drug absorption, we characterized a human small intestinal epithelial cell (HIEC) monolayer that differentiated from adult ISCs. Continuous proliferation/ differentiation from ISCs consistently conferred the capability of maturation of enterocytes to HIECs over 25 passages. The morphologically matured HIEC monolayer consisted of polarized columnar epithelia with dense microvilli, tight junctions, and desmosomes 8 days after seeding onto culture inserts. Transepithelial electrical resistance across the monolayer was 9-fold lower in HIECs (98.9 V 3 cm 2 ) than in Caco-2 cells (900 V 3 cm 2 ), which indicated that the looseness of the tight junctions in the HIEC monolayer was similar to that in the human small intestine (approximately 40 V 3 cm 2 ). No significant differences were observed in the overall gene expression patterns of the major drug-metabolizing enzymes and transporters between the HIEC and Caco-2 cell monolayers. Furthermore, the functions of P-glycoprotein and breast cancer resistance protein in the HIEC monolayer were confirmed by the vectorial transport of marker substrates and their disappearance in the presence of specific inhibitors. The apparent drug permeability values of paracellularly transported compounds (fluorescein isothiocyanate-dextran 4000, atenolol, and terbutaline) and nucleoside transporter substrates (didanosine, ribavirin, and doxifluridine) in the HIEC monolayer were markedly higher than those of Caco-2 cells, whereas transcellularly transported drugs (pindolol and midazolam) were equally well permeated. In conclusion, the HIEC monolayer can serve as a novel and superior alternative to the conventional Caco-2 cell monolayer for predicting oral absorption in humans.

Functional Comparison of Human Colonic Carcinoma Cell Lines and Primary Small Intestinal Epithelial Cells for Investigations of Intestinal Drug Permeability and First-Pass Metabolism

“…At approximately 50 V×cm 2 , TEER values for the fSIEC monolayers were substantially less than that of T84 or Caco-2 but strikingly close to the 40 V×cm 2 reported for human small intestine in an Ussing chamber (Sjoberg et al, 2013). This finding is in contrast to the slightly higher 100 V×cm 2 seen in fSIEC monolayers derived from adult intestinal stems cells or the 200 V×cm 2 seen in those derived from induced pluripotent stem cells (Kauffman et al, 2013;Takenaka et al, 2014). In addition to induced pluripotent stem cell-derived SIEC, Kauffman et al also assessed TEER values in primary human intestinal epithelial cells from an unstated region of the intestine.…”

Characteristic Analysis of Intestinal Transport in Enterocyte-Like Cells Differentiated from Human Induced Pluripotent Stem Cells

“…It was reported that TEER values in the human small intestine were approximately 40 V Â cm 2 (Sjoberg et al, 2013). In addition, we previously reported that the values in human small intestinal epithelial cell (HIEC) monolayer were 98.9 V Â cm 2 , even lower in the Caco-2 cell monolayer (900 VÂcm 2 ) (Takenaka et al, 2014). It was indicated that the TEER value of the enterocyte-like cell membrane was comparable to that of the HIEC monolayer and the human small intestine.…”

“…7). Takenaka et al (2014) reported that the HIEC monolayer with loose tight junctions was able to accurately predict the oral absorption of paracellularly absorbed compounds. The enterocyte-like cell membrane could also be useful for the prediction of the F a values of drugs, including such compounds.…”

Generation of Enterocyte-Like Cells with Pharmacokinetic Functions from Human Induced Pluripotent Stem Cells Using Small-Molecule Compounds

“…CYP3A4 mRNA expression was induced by 1a,25-dihydroxyvitamin D 3 but not by rifampicin (Schmiedlin-Ren et al, 2001;Martin et al, 2008) because the pregnane X receptor (PXR) was not expressed in Caco-2 cells . Recently, a human small intestinal epithelial cell monolayer has been reported to be useful as a novel in vitro system to predict oral absorption in humans (Takenaka et al, 2014). The human small intestinal epithelial cell monolayer expressed various drug transporters and could be used to evaluate the permeability of paracellularly absorbed compounds; however, further exploration of its pharmacokinetic functional characteristics is desirable considering that accurate prediction of intestinal absorption in this cell line is difficult.…”