期刊
DRUG METABOLISM AND DISPOSITION
卷 44, 期 10, 页码 1662-1667出版社
AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS
DOI: 10.1124/dmd.116.069336
关键词
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资金
- Japan Society for the Promotion of Science [23390036, 25860120, 26293036]
- Adaptable & Seamless Technology Transfer Program through Target-Driven R&D (A-STEP) of Japan Science and Technology Agency [AS262Z01122Q]
- Nakatomi Foundation [NF2014-32]
- Grants-in-Aid for Scientific Research [25860120, 15K18929, 23390036, 26293036] Funding Source: KAKEN
We previously demonstrated that differentiated enterocytes from human induced pluripotent stem (iPS) cells exhibited drug-metabolizing activities and cytochrome P450 CYP3A4 inducibility. The aim of this study was to apply human iPS cell-derived enterocytes in pharmacokinetic studies by investigating the characteristics of drug transport into enterocyte-like cells. Human iPS cells cultured on feeder cells were differentiated into endodermal cells using activin A. These endodermal-like cells were then differentiated into intestinal stem cells by fibroblast growth factor 2. Finally, epidermal growth factor and small-molecule compounds induced the maturation of the intestinal stem cell-like cells. After differentiation, we performed transepithelial electrical resistance (TEER) measurements, immunofluorescence staining, and transport studies. TEER values increased in a time-dependent manner and reached approximately 100 Omega x cm(2). Efflux transport of Hoechst 33342, a substrate of breast cancer resistance protein (BCRP), was observed and inhibited by the BCRP inhibitor Ko143. The uptake of peptide transporter 1 substrate glycylsarcosine was also confirmed and suppressed when the temperature was lowered to 4 degrees C. Using immunofluorescence staining, villin and Na+-K+ ATPase were expressed. These results suggest that human iPS cell-derived enterocytes had loose tight junctions, polarity, as well as uptake and efflux transport functions. In addition, the rank order of apparent membrane permeability coefficient (P-app) values of these test compounds across the enterocyte-like cell membrane corresponded to the fraction absorbance (F-a) values. Therefore, differentiated enterocytes from human iPS cells may provide a useful comprehensive evaluation model of drug transport and metabolism in the small intestine.
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