NF-κB Signaling in Ex-Vivo Mouse Intestinal Organoids.

Autor:	Shostak K; Interdisciplinary Cluster for Applied Genoproteomics (GIGA), University of Liege, Liège, Belgium., Wathieu C; Interdisciplinary Cluster for Applied Genoproteomics (GIGA), University of Liege, Liège, Belgium., Tielens S; Interdisciplinary Cluster for Applied Genoproteomics (GIGA), University of Liege, Liège, Belgium., Chariot A; Interdisciplinary Cluster for Applied Genoproteomics (GIGA), University of Liege, Liège, Belgium. Alain.chariot@uliege.be.; Laboratory of Medical Chemistry, GIGA Stem Cells, University of Liège, Liège, Belgium. Alain.chariot@uliege.be.; Walloon Excellence in Life Sciences and Biotechnology (WELBIO), Wavre, Belgium. Alain.chariot@uliege.be.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2366, pp. 283-292.
DOI:	10.1007/978-1-0716-1669-7_17
Abstrakt:	We describe here a protocol to assess NF-κB activation in ex-vivo organoids generated from mouse intestinal crypts. These structures are maintained in culture as crypt-villus forming organoids. These ex-vivo organoids maintain both self-renewal and multilineage differentiation overtime. We also describe the generation of ex-vivo organoids from Apc-mutated mouse intestinal crypts. Both wild-type and Apc-mutated organoids respond very well to NF-κB-activating signals such as TNFα but not to LPS. The kinetic of NF-κB activation in response to these signals in ex-vivo intestinal organoids is very similar to what we see in 2D cell lines. This protocol provides investigators a powerful tool to assess NF-κB activation in both healthy and transformed intestinal epitheliums maintained in culture as 3D structures.
Databáze:	MEDLINE
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