UNIT 10.16 Preparation of Hydrogel Substrates with Tunable Mechanical Properties

  1. Justin R. Tse,
  2. Adam J. Engler

Published Online: 1 JUN 2010

DOI: 10.1002/0471143030.cb1016s47

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Tse, J. R. and Engler, A. J. 2010. Preparation of Hydrogel Substrates with Tunable Mechanical Properties. Current Protocols in Cell Biology. 47:10.16:10.16.1–10.16.16.

Author Information

  1. Department of Bioengineering, University of California, San Diego, La Jolla, California

Publication History

  1. Published Online: 1 JUN 2010
  2. Published Print: JUN 2010


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. Curr. Protoc. Cell Biol. 47:10.16.1-10.16.16. © 2010 by John Wiley & Sons, Inc.


  • extracellular matrix;
  • elasticity;
  • hydrogel;
  • gradient;
  • polyacrylamide