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

Biol Open. 2020; 
Kondo S, Mizuno S, Hashita T, Iwao T, Matsunaga T
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Recombinant Proteins Materials were purchased from the following sources: fibroblast growth factor (FGF) 2 and activin A (PeproTech, Rocky Hill, NJ, USA); iMatrix-511 (Nippi, Tokyo, Japan); CHIR99021 and Y-27632 (Focus Biomolecules, Plymouth Meeting, PA, USA); KSR, DMEM, DMEM/F-12, Advanced DMEM/F-12, N2 supplement and B27 serum-free supplement (Thermo Fisher Scientific, Waltham, MA, USA); FBS (Nichirei Biosciences, Tokyo, Japan); R-spondin 1, Noggin and epidermal growth factor (EGF) (GenScript, Piscataway, NJ, USA); N-acetylcysteine (Sigma-Aldrich, St Louis, MO, USA); 8-Br-cAMP (Enzo Life Sciences, New York, NY, USA); PD98059 and A-83-01 (AdooQ BIOSCIENCE, Irvine, CA, USA); 5-aza-2′-deoxycytidine (Chem-impex International, Wood Dale, IL, USA); L-glutamine, minimum essential medium nonessential amino acid solution (NEAA), penicillin-streptomycin solution, valproic acid and nicotinamide (Wako Pure Chemical Industries, Osaka, Japan); and total RNA from human small intestine samples (5 donors; BioChain Institute, Newark, CA, USA). All other reagents were of the highest quality available, including SB 202190 (ChemScene, Monmouth Junction, NJ, USA) and WNT3a (ATGen Co. Ltd., Gyeonggi-do, South Korea). Get A Quote

摘要

The small intestine plays an important role in the pharmacokinetics of orally administered drugs due to the presence of drug transporters and drug-metabolizing enzymes. However, few appropriate methods exist to investigate intestinal pharmacokinetics. Induced pluripotent stem (iPS) cells can form various types of cells and represent a potentially useful tool for drug discovery. We previously reported that differentiated enterocytes from human iPS cells are useful for pharmacokinetic studies; however, the process is time and resource intensive. Here, we established a new two-dimensional culture method for maintaining human iPS-cell-derived intestinal stem cells (ISCs) with differentiation potency and evaluated t... More

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