Our research goal is to establish robust techniques to generate staple intestinal monolayer interfaces derived from adult bovine small and large intestinal organoids. We employed a multimodal analytical approach to confirm the functional epithelial barrier formation within the established monolayers. Unlike three-dimensional models, 2D monolayers offer an exposed, accessible luminal surface crucial for studying host pathogen interactions.
These monolayers reflecting the in vivo gut with multiple cell lineages now provide tailored protocols for investigating pathogens in specific gut sections such as small and large intestinal niches. Our research identified optimized culture conditions to create and sustain 2D monolayers from cells of bovine mall and large intestinal organoids. We confirmed the formation of functional epithelial barrier within these monolayers using transepithelial electrical registrants and paracellular permeability assays, as well as immunocytochemistry staining techniques.
Our protocol addressed the research gap by focusing on the lack of intestinal organoid derived monolayer models of bovines. These models have been well studied in mice and humans, however, bovines, which are major reservoirs for intestinal pathogens with serious public health implications haven't been adequately explored. Our research establishes and characterizes a 2D bovine monolayer culture system.
This innovative tool allows for the exploration of normal and disease states of bovine intestinal physiology, with potential applications extending into biomedical and translational research of public health significance.
