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Hydrogel-based microchannels to measure confinement- and stiffness-sensitive Yes-associated-protein activity in epithelial clusters

Published online by Cambridge University Press:  07 September 2017

Samila Nasrollahi  and
Amit Pathak Open the ORCID record for Amit Pathak [Opens in a new window]
Samila Nasrollahi
Affiliation:
Department of Mechanical Engineering and Materials Science, Washington University, Saint Louis, MO 63130, USA
Amit Pathak*
Affiliation:
Department of Mechanical Engineering and Materials Science, Washington University, Saint Louis, MO 63130, USA
*
Address all correspondence to Amit Pathak at [email protected]
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Abstract

Nuclear translocation of Yes-associated-protein (YAP) in single cells serves as a key sensor of matrix stiffness. On two-dimensional (2D) polyacrylamide (PA) hydrogels, we found that nuclear YAP localization in epithelial clusters increases with gel stiffness and reduces with cell density. To measure YAP activity in 3D-like confinement of tunable stiffness, we fabricated PA-based microchannels. Here, narrower channels enhanced nuclear YAP localization even in softer extracellular matrix and denser epithelial clusters, both of which reduced YAP activation in 2D. Thus, the presented hydrogel microchannel-based platform may reveal new mechanosensitive cellular signatures in 3D-like settings, which cannot be captured on standard 2D hydrogels.

Type
Biomaterials for 3D Cell Biology Research Letters
Information
MRS Communications , Volume 7 , Issue 3 , September 2017 , pp. 450 - 457
Copyright
Copyright © Materials Research Society 2017 

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