Porcine enteroids mimic the response of the small intestine to ETEC-derived enterotoxins. Spheroids derived from duodenum, jejunum and ileum 6 days after passaging were stimulated with enterotoxins or guanylin and imaged using live-cell microscopy. The surface area of the spheroids was measured using ImageJ. A Representative images displaying ileal spheroid swelling induced by guanylin (10 µM) at T0, T50 and T110 upon administration. B, C The average relative area increase of the spheroids was plotted in function of the time after enterotoxin administration. (n = 3 for all tissues). D Spheroid bursting upon guanylin (10 µM) stimulation. Images are representative for other tissues and swelling inducers. Scale bar = 100 µm. Relative IL8 secretion in medium supernatant (E) and Matrigel dome (F) of jejunal enteroids stimulated for 24 h with bacterial supernatant with (WT) or without enterotoxins (toxin negative) compared to non-immunogenic guanylin (n = 3; Kruskal–Wallis test).