Preparation of Hydrogel Substrates with Tunable Mechanical Properties
Abstract
The modulus of elasticity of the extracellular matrix (ECM), often referred to in a biological context as "stiffness," naturally varies within the body, e.g., hard bones and soft tissue. Moreover, it has been found to have a profound effect on the behavior of anchorage-dependent cells. The fabrication of matrix substrates with a defined modulus of elasticity can be a useful technique to study the interactions of cells with their biophysical microenvironment. Matrix substrates composed of polyacrylamide hydrogels have an easily quantifiable elasticity that can be changed by adjusting the relative concentrations of its monomer, acrylamide, and cross-linker, bis-acrylamide. In this unit, we detail a protocol for the fabrication of statically compliant and radial-gradient polyacrylamide hydrogels, as well as the functionalization of these hydrogels with ECM proteins for cell culture. Included as well are suggestions to optimize this protocol to the choice of cell type or stiffness with a table of relative bis-acrylamide and acrylamide concentrations and expected elasticity after polymerization.
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Pore sizes were in the ideal range of 200-400 μm, known to facilitate osteo-differentiation and functional bone formation.
“Pore sizes were in the ideal range of 200-400 μm, known to facilitate osteo-differentiation and functional bone formation [12,57,58].”
From <i>In vitro</i> degradation and mechanical properties of PLA-PCL copolymer unit cell scaffolds generated by two-photon polymerization (2016) · cites this paper as [12]
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- Authors
- Tse, J. R., & Engler, A. J.
- Affiliations
- University of California San Diego (US)
- Venue
- Current Protocols in Cell Biology
- Publisher
- Wiley
- Published
- 2010-06
- Volume
- 47
- Issue
- 1
- Pages
- Unit 10.16-Unit 10.16
- ISSN
- 1934-2500 (ISSN-L)
- 1934-2616
- OpenAlex
- W1587510658
- Cited by
- 1,070
- References
- 3
- Citations by year
- 2026: 60 · 2025: 104 · 2024: 94 · 2023: 94 · 2022: 88 · 2021: 91 · 2020: 85 · 2019: 83 · 2018: 84 · 2017: 62 · 2016: 57 · 2015: 58 · 2014: 45 · 2013: 33 · 2012: 17
- FWCI
- 8.95
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Current Protocols in Cell Biology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- polyacrylamide hydrogel, hydrogel stiffness, extracellular matrix stiffness, mechanical properties, static compliance, acrylamide, hydrogel functionalization, cell-matrix interactions, adherent cells
- MeSH terms
- Animals, Biomechanical Phenomena, Chemistry Techniques, Analytical, Cross-Linking Reagents, Extracellular Matrix, Humans, Models, Biological, Photochemistry, Hydrogels, Acrylamide
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