6 Gy gamma-irradiation of mouse small intestinal organoids at 24h, 48h and 96h after irradiation

The intact intestinal epithelial barrier protects our body from a range of immune mediated diseases. The epithelial layer has an impressive ability to reconstitute and repair upon damage and this process of repair is increasingly seen as a therapeutic target. In vitro models to study this process in primary intestinal cells are lacking. Therefore we established and characterized an in vitro model of intestinal damage and repair by applying γ-radiation on small intestinal organoids. We then used this model to identify novel regulators of intestinal regeneration. Small intestinal single crypts were isolated from C57BL/6 mice and cultured in matrigel. We treated the organoid cultures, 3 days after seeding, with 6 Gy of gamma-irradiation. RNA was isolated at 24, 48 and 96 hours after irradiation. Non-irradiated controls were collected per time point. 4 replicates per condition. Each time point has its own control.