Red panda intestinal organoids for the functional analysis of Lactobacillus salivarius

Organoids are excellent and reproducible <i>in vitro</i> models for studying physiological mechanisms and modeling diseases in endangered species. However, intestinal organoids of red pandas have not yet been established. This study reports a culture system of intestinal stem cell-derived organoids of red panda that can be passaged, cryopreserved, and resuscitated <i>in vitro</i>. The single-cell RNA sequencing and RNA sequencing analysis showed that red panda intestinal organoids retain tissue-specific cell types and functions. In order to verify the application potential of <i>Lactobacillus salivarius</i> P103 as a probiotic in red pandas, these organoids were used to establish a co-culture system with <i>L</i>. <i>salivarius</i> P103. The results showed that <i>L. salivarius</i> P103 could promote mucin secretion, reduce intestinal inflammation, and inhibit infection induced by <i>Klebsiella pneumoniae</i> 28. The establishment of red panda intestinal organoids provides a long-term, reproducible model system for studying host-microbe interaction mechanisms and decreasing dependence on animal models in wildlife research.