Hydrogel Micropost Arrays with Single Post Tunability to Study Cell Volume and Mechanotransduction

The extracellular matrix varies considerably in mechanical properties at the microscale. It remains unclear how cells respond to these properties, in part, due to lack of tools to create precisely defined microenvironments in a discrete manner. Here, freeform stereolithography is leveraged to control the placement and elastic modulus of individual hydrogel microposts that serve as discrete matrix signals to interface with cells. Mesenchymal stromal cells (MSCs) located in the interstitial spaces between microposts above a base layer are analyzed. Cell volume is higher when MSCs interact with more microposts. MSCs show higher strain energy when they interact simultaneously with 4-kPa and 20-kPa microposts than with mechanically homogeneous micropost arrays. MSCs are sensitive to pharmacological inhibition of Rho-associated protein kinase in 4-kPa arrays, but resistant when presented together with 20-kPa arrays. Yes-associated protein (YAP) activity increases with higher cell volume and elastic modulus of microposts. Surprisingly, YAP activity becomes less variable with higher cell volume and decreases with higher average force and strain energy per post when MSCs interact with both 4-kPa and 20-kPa microposts simultaneously. Together, these results describe a material system for systematically investigating how the placement and intrinsic properties of discrete matrix signals impact cell volume and mechanotransduction.