Large polyacrylamide hydrogels for large-batch cell culture and mechanobiological studies

The rigidity of a cell’s substrate or extracellular matrix plays a vital role in regulating cell and tissue functions. Polyacrylamide (PAAm) hydrogels are one of the most widely used cell culture substrates that provide a physiologically relevant range of stiffness. However, it is still arduous and time-consuming to prepare PAAm substrates in large batches for high-yield or multi-scale cell cultures. Here we present a simple method to prepare PAAm hydrogels with less time cost and easily accessible materials. The hydrogel is mechanically uniform and supports cell culture in a large batch. We further show that the stiffness of the hydrogel covers a large range of Young’s modulus and is sensed by cells, regulating various cell features including changes in cell morphology, proliferation, and contractility. This method improves reproducibility of mechanobiology studies and can be easily applied for mechanobiology research requiring large numbers of cells or experimental groups.