Arthur G. James Cancer Hospital and Richard J. Solove Research Institute 1 article published in JoVE Bioengineering Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels Abdulaziz S. Fakhouri1,2,3, Jennifer L. Leight1,2 1Department of Biomedical Engineering, The Ohio State University, 2The Ohio State University Comprehensive Cancer Center, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, 3Biomedical Technology Department, King Saud University Here, a protocol is presented for encapsulating and culturing cells in poly(ethylene glycol) (PEG) hydrogels functionalized with a fluorogenic matrix metalloproteinase (MMP)-degradable peptide. Cellular MMP and metabolic activity are measured directly from the hydrogel cultures using a standard microplate reader.