Developing an advanced gut on chip model enabling the study of epithelial cell/fibroblast interactions

Organoids are widely used as a model system to study gut pathophysiology; however, they fail to fully reproduce the complex, multi-component structure of the intestinal wall. We present here a new gut on chip model that allows the co-culture of primary epithelial and stromal cells. The device has the topography and dimensions of the mouse gut and is based on a 3D collagen I scaffold. The scaffold is coated with a thin layer of laminin to mimic the basement membrane. To maintain the scaffold structure while preserving its cytocompatibility, the collagen scaffold was rigidified by threose-based post-polymerization treatment. This treatment being cytocompatible enabled the incorporation of primary intestinal fibroblasts inside the scaffold, reproducing the gut stromal compartment. We observed that mouse organoids, when deposited into crypts, opened up and epithelialized the scaffold, generating a polarized epithelial monolayer. Proper segregation of dividing and differentiated cells along the crypt-villus axis was achieved under these conditions. Finally, we show that the application of fluid shear stress allows the long-term culture of this intestinal epithelium. Our device represents a new biomimetic tool that captures key features of the gut complexity and could be used to study gut pathophysiology.

Automated summary

A new gut on chip model has been developed for co-culture of primary epithelial and stromal cells, mimicking the structure and topography of the mouse gut. The collagen scaffold is rigidified by threose-based post-polymerization treatment to maintain its structure and cytocompatibility. Mouse organoids deposited into crypts are able to epithelialize the scaffold and form a polarized epithelial monolayer, demonstrating proper cell segregation along the crypt-villus axis. Fluid shear stress application allows long-term culture of the intestinal epithelium in this biomimetic tool.