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Open Access Publications from the University of California

Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels.

  • Author(s): Hadden, William J
  • Young, Jennifer L
  • Holle, Andrew W
  • McFetridge, Meg L
  • Kim, Du Yong
  • Wijesinghe, Philip
  • Taylor-Weiner, Hermes
  • Wen, Jessica H
  • Lee, Andrew R
  • Bieback, Karen
  • Vo, Ba-Ngu
  • Sampson, David D
  • Kennedy, Brendan F
  • Spatz, Joachim P
  • Engler, Adam J
  • Choi, Yu Suk
  • et al.

The spatial presentation of mechanical information is a key parameter for cell behavior. We have developed a method of polymerization control in which the differential diffusion distance of unreacted cross-linker and monomer into a prepolymerized hydrogel sink results in a tunable stiffness gradient at the cell-matrix interface. This simple, low-cost, robust method was used to produce polyacrylamide hydrogels with stiffness gradients of 0.5, 1.7, 2.9, 4.5, 6.8, and 8.2 kPa/mm, spanning the in vivo physiological and pathological mechanical landscape. Importantly, three of these gradients were found to be nondurotactic for human adipose-derived stem cells (hASCs), allowing the presentation of a continuous range of stiffnesses in a single well without the confounding effect of differential cell migration. Using these nondurotactic gradient gels, stiffness-dependent hASC morphology, migration, and differentiation were studied. Finally, the mechanosensitive proteins YAP, Lamin A/C, Lamin B, MRTF-A, and MRTF-B were analyzed on these gradients, providing higher-resolution data on stiffness-dependent expression and localization.

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