Video 2_Development of a novel organoid-based two-dimensional system to model the intestinal host–parasite interface in fasciolosis.avi

Here, we establish a human duodenum organoid-derived epithelial monolayer (ODM) co-culture system to investigate the early host–parasite interface, providing a controlled experimental platform for analysing interactions between FhNEJ and the intestinal epithelium.</p>Methods<p>ODMs representing the human duodenum were co-cultured with either live FhNEJ or a tegument-enriched protein fraction derived from the parasites.

The ODM co-culture system was subsequently employed to assess FhNEJ viability and parasite–epithelial interactions together with complementary functional and molecular read-outs. These included measurements of trans-epithelial electrical resistance, fluorescein permeability assays, immunofluorescence analysis of intercellular epithelial junctions, and quantitative proteomic profiling.</p>Results<p>The ODM–FhNEJ co-culture system supported sustained parasite viability and enabled direct visualisation of intimate parasite–epithelial interactions.