Biological and Mechanical Influence of Three-Dimensional Microenvironment Formed in Microwell on Multicellular Spheroids Composed of Heterogeneous Hair Follicle Stem Cells

This study investigates the effects of a three-dimensional (3D) microenvironment on hair follicle (HF) regeneration by culturing HF-derived cells, primarily hair follicle stem cells (HFSCs), in 3D spheroids using a microwell array. The research focuses on the expression of Sonic hedgehog signaling and stem cell markers in these spheroids, as well as the impact of microwell depth on spheroid formation and biological conditions. Computational fluid dynamics were used to predict biomolecular diffusion and convective flow, providing insights into the physical stimulations at the micro-scale. The findings offer fundamental insights into HF physiology in a 3D environment and highlight the importance of geometric parameters in microwell systems for HF research.