Establishment of a Porcine Small Intestinal Organoid Model for Porcine Rotavirus Infection.

Porcine rotavirus (PoRV), a major enteric pathogen, causes severe diarrhea, dehydration, high morbidity, and mortality, leading to significant economic losses in the porcine industry. In this study, we established a porcine small intestinal organoid culture system and developed an in vitro model for PoRV infection. Jejunal crypts were isolated from neonatal piglets and subsequently cultured to generate porcine small intestinal organoids in both three-dimensional (3D) and two-dimensional (2D) organoid formats, properties of which were verified via morphological assessment and immunostaining for Lgr5, villin, Ki-67, and lysozyme. Furthermore, a PoRV infection model was established in both 3D and 2D organoid cultures, and growth kinetics demonstrated efficient PoRV replication. PoRV infection also significantly increased the expression of inflammatory cytokines, including IL-6 and IFN-α. Collectively, these results demonstrate that the established porcine small intestinal organoid models support efficient PoRV replication and provide a physiologically relevant platform for studying PoRV pathogenesis and evaluating potential antiviral strategies.