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Much of what we know about the early stages of T cell activation has been obtained from studies of T cells interacting with glass-supported lipid bilayers that favor imaging but are orders of magnitude stiffer than typical cells. We developed a method for attaching lipid bilayers to polydimethylsiloxane polymer supports, producing "soft bilayers" with physiological levels of mechanical resistance (Young's modulus of 4 kPa). Comparisons of T cell behavior on soft and glass-supported bilayers revealed that whereas late stages of T cell activation are thought to be substrate-stiffness dependent, early calcium signaling was unaffected by substrate rigidity, implying that early steps in T cell receptor triggering are not mechanosensitive. The exclusion of large receptor-type phosphatases was observed on the soft bilayers, however, even though it is yet to be demonstrated at authentic cell-cell contacts. This work sets the stage for an imaging-based exploration of receptor signaling under conditions closely mimicking physiological cell-cell contact. Copyright © 2020 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Citation

Anna H Lippert, Ivan B Dimov, Alexander K Winkel, Jane Humphrey, James McColl, Kevin Y Chen, Ana M Santos, Edward Jenkins, Kristian Franze, Simon J Davis, David Klenerman. Soft Polydimethylsiloxane-Supported Lipid Bilayers for Studying T Cell Interactions. Biophysical journal. 2021 Jan 05;120(1):35-45

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PMID: 33248128

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