The extracellular matrix plays a major role in defining the microenvironment of cells and in modulating cell behavior and phenotype. We describe a rapid method for the isolation of cell-derived extracellular matrix, which can be adapted to different scales for microscopic, biochemical, proteomic, or functional studies.
Total time
~2–4 hours per sample (depending on scale and downstream analysis)
Model organism
Cell culture (fibroblasts and other cell types)
Steps
1
Remove cells with ammonium hydroxide solution
Treat cell-seeded surfaces with ammonium hydroxide to selectively lyse and remove cellular material while preserving the underlying extracellular matrix.
▶ 00:56
2
Prepare sterile ECM for functional assays
Process decellularized matrix under sterile conditions and prepare it for use in cell functional studies and behavior assays.
▶ 03:21
3
Isolate ECM for biochemical and proteomic analysis
Collect and process the isolated extracellular matrix for protein extraction, SDS-PAGE, immunoblotting, and mass spectrometry-based proteomics.
▶ 05:01
4
Analyze isolated ECM composition and structure
Characterize the recovered extracellular matrix using microscopy, biochemical assays, and molecular profiling to confirm successful isolation and assess matrix properties.
▶ 06:30