A human intestinal epithelial-mesenchyme-immune triple culture system for disease modelling

The intestinal mucosa plays a vital role in nutrient absorption, drug metabolism, and pathogen defence. Advances in single-cell technologies have highlighted the specialised roles of various cell types that execute these diverse functions. Beyond intestinal epithelial cells, fibroblasts regulate the extracellular matrix (ECM) and modulate pro-inflammatory signalling, while antigen-presenting cells (macrophages and dendritic cells) maintain intestinal homeostasis and immune responses. However, the incorporation of such cellular complexity within existing in vitro models of the human intestine is currently lacking.

We developed a human intestinal co-culture model that incrementally mimics the mucosal cellular environment, comprising intestinal epithelial cells, intestinal fibroblasts, and antigen-presenting cells. The model incorporated co-cultures of both adult and foetal cells to facilitate studies of intestinal development, barrier function, inflammation, and viral infections.

We successfully established an advanced multi-cellular intestinal co-culture model that recapitulates key features of the native mucosal environment. This model demonstrated the capacity for ECM deposition, Paneth cell differentiation, and functional immune interactions. Notably, the co-culture system effectively modelled immune responses during viral infections, demonstrating the utility of incorporating multiple cell types and developmental stages.

Our co-culture model improves the physiological relevance of in vitro studies by incorporating multi-cellular complexity previously lacking in existing systems. This advancement enables the exploration of epithelial-mesenchymal-immune crosstalk in intestinal health and disease, providing a powerful platform for studying intestinal development, barrier dysfunction, and infection-related pathology.