TURNING MICRO-WELLS INTO ARTIFICIAL 3D MICRO-NICHES WITH COMBINED BIOCHEMICAL AND BIOPHYSICAL CUES

Abstract

Cells in a tissue are residing in a complex and three-dimensional (3D) micro-environment that governs their behavior. While a large number of environmental cues can be reconstituted individually and mostly in two-dimension, it remains challenging to control in a single technique any combination of these factors in-vitro. In the first part of the thesis, we propose a novel approach to create 3D artificial micro-niches with more than 80 possible combinations of environmental factors, bio-functionalizing micro-wells simultaneously with protein patterns, structured hydrogels and topographical features with micrometer precision. The second part is dedicated to cell seeding and behavior inside these complex micro-niches, focusing on the polarization of single epithelial cell. In the last part, we investigate the potential of our method at the organoid scale. To explore this topic, we study the epithelial-mesenchymal interaction necessary for hair growth by monitoring the spatial organization of human hair follicle cells inside the micro-niches.