Development of an ex vivo intestinal GVHD model using human intestinal organoids and peripheral T cells. (A) Representative images of human small intestinal organoids cocultured for 8 hours with syngeneic (syn) or allogeneic (allo) human T cells. Sorted T cells were stained with CellBrite Green (green) before coculture, and PI (red) was added into the culture medium at the beginning to stain dead organoids. Scale bars, 25 µm. Viability of human small intestinal organoids from 20 patients (supplemental Table 3) at 48 hours after stimulation with 50 ng/mL TNF-α or post coculture with allogeneic and syngeneic T cells (B) or with only allogeneic T cells (C). (D) Viability of human colonic organoids from 4 patients (supplemental Table 2) at 48 hours after stimulation with 50 ng/mL TNF-α or post coculture with allogeneic and/or syngeneic T cells. At least 2 independent experiments were performed. *P < .05, **P < .01, ***P < .001, ****P < .0001. Pt., patient.