Towards 3D Traction Force Microscopy of Intestinal Organoids

Intestinal organoids mimic key features of the intestinal epithelium, from 3D shape to crypt curvature and cellular function and compartmentalization. All of these characteristics are closely linked to the internal stresses of the epithelium and the traction forces exerted on their environment. Recent studies have shown the role of those traction stresses in homeostasis of open-lumen flattened intestinal organoids, where organoids are broken into pieces and seeded on top of a flat hydrogel. However, in this system the intestinal organoids lose their characteristic 3D structure and curvature of their different regions, that we believe to be key in the study of epithelial function. In this project, we aim to develop a novel platform to measure the 3D traction forces exerted by open-lumen 3D intestinal organoids, when cultured on a physiologically shaped crypt-villi scaffold. It will involve microfabrication techniques to manufacture the required 3D shaped elastic hydrogels, advanced cell culture techniques to attach the organoids on the hydrogel and promote the correct placement of the different cell types, microscope imaging to characterize the system and measure the hydrogel deformation and computer analysis to measure the exerted tractions from the measured gel deformations.