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Published on: September 18, 2016
Three-Dimensional High-Throughput Cell Encapsulation Platform to Study Changes in Cell-Matrix Interactions.
Kelly M Mabry1, Megan E Schroeder1, Samuel Z Payne1
1Department of Chemical and Biological Engineering, ‡Department of Materials Science, and §Howard Hughes Medical Institute and the BioFrontiers Institute, University of Colorado at Boulder , Boulder, Colorado 80303, United States.
Researchers developed a high-throughput platform to study cell-matrix interactions using peptide-functionalized hydrogels. This method dynamically alters biochemical and mechanical cues, revealing how fibronectin peptides influence cell behavior and myofibroblast activation in aortic valvular interstitial cells.
Area of Science:
- Biomaterials Science
- Cell Biology
- Tissue Engineering
Background:
- Cells in their native environment sense complex biochemical and mechanical signals.
- High-throughput screening of cell-laden matrices is crucial for understanding cell-ECM signaling.
- Aortic valvular interstitial cells (VICs) play a key role in aortic stenosis pathogenesis.
Purpose of the Study:
- To develop a high-throughput platform for encapsulating cells in dynamically tunable hydrogels.
- To investigate the impact of time-varying matrix cues on VIC behavior.
- To characterize VIC response to adhesive peptide (RGDS) and matrix stiffness.
Main Methods:
- Fabrication of peptide-functionalized poly(ethylene glycol) hydrogels via thiol-ene photoclick reaction.
- In situ modification of hydrogel properties: RGDS peptide tethering (0-1.5 mM) and matrix modulus alteration (1-6 kPa).
- Assessment of VIC morphology and α-smooth muscle actin (αSMA) stress fiber expression to quantify myofibroblast activation.
Main Results:
- VICs exhibited significant elongation upon RGDS addition within 24 hours.
- Myofibroblast activation in VICs was dependent on RGDS presence, particularly in 1 kPa gels (16-24% activation).
- VIC morphology and activation showed path-dependence, influenced by exposure time to RGDS and matrix stiffness.
Conclusions:
- The developed platform enables real-time modification of the cell microenvironment for high-throughput screening.
- Dynamic changes in matrix stiffness and adhesion significantly impact VIC morphology and myofibroblast activation.
- Understanding these cell-matrix interactions is vital for developing therapies for fibrotic diseases like aortic stenosis.
