We have developed a technology for maintaining muscle stem cell (MuSC) regenerative potential in culture by reconstructing the biomechanical environment of its in vivo niche. This enables us to perform time-course studies of MuSCs to elucidate their fate determinants. It also allows us to expand them and treat them to enhance their therapeutic capacity.
Hydrogels are inert to protein adsorption and mimic natural tissue in that they are soft and highly hydrated (up to 98% water), thus allowing diffusion of nutrients and presentation of proteins in a more physiological manner than conventional plastic dishes. Using this innovative platform we can control protein identity, concentration, and presentation, as well as niche size, and hydrogel matrix mechanical properties. The hydrogel substrates are completely transparent, providing excellent optics for microscopy. Microwell arrays make it possible to track cell fate changes via time-lapse microscopy for clonal populations of single stem cells and perform quantitative statistical analyses of outcomes.