High-Throughput Three-Dimensional Hydrogel Cell Encapsulation Assay for Measuring Matrix Metalloproteinase Activity

Area of Science:

• Biotechnology
• Cell Biology
• Drug Discovery

Background:

• Three-dimensional (3D) cell cultures better replicate in vivo conditions than 2D cultures.
• Current high-throughput (HT) 3D assays are primarily limited to cell viability measurements.
• There is a need for HT assays measuring diverse cellular functions for drug screening.

Purpose of the Study:

• To adapt a hydrogel system for simultaneous detection of matrix metalloproteinase (MMP) and metabolic activity in 3D cell cultures.
• To validate this system for high-throughput (HT) drug screening applications.

Main Methods:

• Cells were encapsulated in a hydrogel for 3D culture.
• Simultaneous detection of matrix metalloproteinase (MMP) and metabolic activity was enabled.
• Assay performance was characterized using Z'-factor, coefficient of variation, and drug treatment of fibrosarcoma cells.

Main Results:

• The assay achieved high Z'-factor values (>0.9 enzymatic, >0.5 cellular) and low intraplate coefficients of variation (<10% enzymatic, <12% cellular).
• Dimethyl sulfoxide did not affect signal measurement.
• The assay demonstrated robust signals with various cell types and successfully detected drug effects on MMP activity.

Conclusions:

• This novel hydrogel-based assay enables simultaneous HT measurement of MMP and metabolic activity in 3D cell cultures.
• The system is suitable for drug screening and development, offering a more comprehensive analysis of cellular responses.
• It addresses the need for advanced HT 3D assays beyond simple cell viability measurements.