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Gastrointestinal Epithelial Organoid Cultures from Postsurgical Tissues.

Methods in molecular biology (Clifton, N.J.) (2017-08-18)
Soojung Hahn, Jongman Yoo
ZUSAMMENFASSUNG

An organoid is a cellular structure three-dimensionally (3D) cultured from self-organizing stem cells in vitro, which has a cell population, architectures, and organ specific functions like the originating organs. Recent advances in the 3D culture of isolated intestinal crypts or gastric glands have enabled the generation of human gastrointestinal epithelial organoids. Gastrointestinal organoids recapitulate the human in vivo physiology because of all the intestinal epithelial cell types that differentiated and proliferated from tissue resident stem cells. Thus far, gastrointestinal organoids have been extensively used for generating gastrointestinal disease models. This protocol describes the method of isolating a gland or crypt using stomach or colon tissue after surgery and establishing them into gastroids or colonoids.

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Albumin aus Rinderserum, lyophilized powder, ≥96% (agarose gel electrophoresis)
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N-Acetyl-L-Cystein, BioReagent, suitable for cell culture
Sigma-Aldrich
Y-27632 -dihydrochlorid, ≥98% (HPLC)
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CHIR99021, ≥98% (HPLC)
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Ethylendiamintetraessigsäure, ACS reagent, 99.4-100.6%, powder
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SB 202190, ≥98% (HPLC)
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A 83-01, ≥98% (HPLC)
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[Leu15]-Gastrin I Human, ≥95% (HPLC)
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Fetales Kälberserum, Australia origin, USDA approved, sterile-filtered, suitable for cell culture
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Thiazovivin, ≥98% (HPLC)