Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels

3D culture makes it more difficult to measure cellular function, using standard biological assays. This protocol enables measurement of cellular and somatic activity without further sample processing, using a fluorescent sensor and a standard microplate reader. This protocol enables a number of new applications, including high throughput drug screening.

Additionally, the fluorescent sensor can be exchanged with a different sensor to measure other proteases or cell functions. It is important to carefully plan out the experiment in advance, complete all calculations for the hydrogel preparation, and setup all equipments and reagents to minimize the time cells are in suspension. To begin, prepare the assay media with 1%charcoal stripped Fetal Bovine Serum, two mM L-glutamine, 10 units per mL of penicillin, and 10 micrograms per mL of streptomycin.