A SILAC-Based Method for Quantitative Proteomic Analysis of Intestinal Organoids.

ABSTRACT: Organoids have the potential to bridge 3D cell culture to tissue physiology by providing a model resembling in vivo organs. Long-term growing organoids were first isolated from intestinal crypt cells and recreated the renewing intestinal epithelial niche. Since then, this technical breakthrough was applied to many other organs, including prostate, liver, kidney and pancreas. We describe here how to apply a SILAC-based quantitative proteomic approach to measure protein expression changes in intestinal organoids under different experimental conditions. We generated SILAC organoid media that allow organoids to grow and differentiate normally, and confirmed the incorporation of isotopically labelled amino acids. Furthermore, we used a treatment reported to affect organoid differentiation to de

SUBMITTER: Gonneaud A 

PROVIDER: S-EPMC5128881 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature