3 years ago

Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels [Cell Biology]

Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels [Cell Biology]
Yu Suk Choi, Jennifer L. Young, Du Yong Kim, Joachim P. Spatz, Hermes Taylor-Weiner, Jessica H. Wen, Karen Bieback, Andrew R. Lee, Andrew W. Holle, Philip Wijesinghe, David D. Sampson, Ba-Ngu Vo, William J. Hadden, Brendan F. Kennedy, Adam J. Engler, Meg L. McFetridge

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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