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A Rapid, Scalable Method for the Isolation, Functional Study, and Analysis of Cell-derived Extracellular Matrix
Published on: January 4, 2017
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A Rapid, Scalable Method for the Isolation, Functional Study, and Analysis of Cell-derived Extracellular Matrix
Andrew L Hellewell1, Silvia Rosini1, Josephine C Adams2
1School of Biochemistry, University of Bristol.
Journal of Visualized Experiments : Jove
|January 25, 2017
Summary
Researchers developed a rapid method to isolate cell-derived extracellular matrix (ECM) for experiments. This technique simplifies studying ECM composition and dynamics in various cell types.
Area of Science:
- Biochemistry
- Cell Biology
- Biomaterials Science
Background:
- The extracellular matrix (ECM) is crucial in physiological and pathological processes.
- Isolating ECM is challenging due to its complex, insoluble nature and potential cellular contamination.
Purpose of the Study:
- To present a rapid and reliable method for isolating cell-derived ECM from cultured cells.
- To enable downstream experimentation, including visualization, biochemical analysis, and proteomics.
Main Methods:
- A novel method for rapid cell removal from cultured cells to isolate the underlying ECM.
- Utilizes techniques like in situ immunofluorescence microscopy, SDS-PAGE, immunoblotting, and mass spectrometry.
- Allows for sterile ECM isolation for functional studies.
Main Results:
- Successfully isolated cell-derived ECM, enabling visualization of ECM components.
- Facilitated tracking of ECM protein dynamics using fluorescence microscopy.
- Enabled comprehensive biochemical and proteomic analyses of the isolated ECM.
Conclusions:
- The described method provides a straightforward approach to isolate and analyze cell-derived ECM.
- This technique supports diverse experimental designs, including functional studies with sterile ECM layers.
- It offers a valuable tool for investigating ECM's role in cellular processes.
