Dmd059493 1947..1954

نویسندگان

  • Toru Takenaka
  • Naomoto Harada
  • Jiro Kuze
  • Masato Chiba
  • Takahiro Iwao
  • Tamihide Matsunaga
چکیده

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) than in Caco-2 cells (900 V 3 cm), which indicated that the looseness of the tight junctions in the HIECmonolayer was similar to that in the human small intestine (approximately 40 V 3 cm). 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

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Krebs, H. A. & Henseleit, K. (1932). Hoppe-Seyl. Z. 210,33. Liebecq, C. (1953). Arch. int. Phy8iol. 61, 550. Long, C. (1952). Biochem. J. 50, 407. Mentha, J. & Voegtli, W. (1947). Helv. phy8iol. acta, 5, C43. Nelson, N. (1944). J. biol. Chem. 153, 375. Perlmutter, M., Weisenfeld, S. & Mufson, M. (1952). Endocrinology, 50, 442. Riesser, 0. (1947). Biochim. biophys. Acta, 1, 208. Snedecor, G. M. ...

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تاریخ انتشار 2014