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  • Gastrointestinal Epithelial Organoid Cultures from Postsurgical Tissues.

Gastrointestinal Epithelial Organoid Cultures from Postsurgical Tissues.

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

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
SB 202190, ≥98% (HPLC)
Sigma-Aldrich
Y-27632 dihydrochloride, ≥98% (HPLC)
Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Sigma-Aldrich
Fetal Bovine Serum, Australia origin, USDA approved, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
N-Acetyl-L-cysteine, BioReagent, suitable for cell culture
Sigma-Aldrich
A 83-01, ≥98% (HPLC)
Sigma-Aldrich
Thiazovivin, ≥98% (HPLC)