PDMS Hydrogel-coated Tissue Culture Plates for Studying the Impact of Substrate Stiffness on Dendritic Cell Function

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2022 Mar 22

PMID 35313712

Authors

Megan Lee

Kevin Chu

Mainak Chakraborty

Nicholas Kotoulas

Masoud Akbari

Cynthia Goh

Xavier Clemente-Casares

Daniel A Winer

Annie Shrestha

Sue Tsai

Affiliations

Soon will be listed here.

Abstract

The mechanical properties of polydimethylsiloxane hydrogels can be tuned to mimic physiological tensions, an underappreciated environmental parameter in immunology studies. We describe a workflow to prepare PDMS-coated tissue culture plates with biologically relevant substrate stiffness, and the use of these hydrogel plates to condition isolated primary splenic CD11c+ dendritic cells (DC). Finally, we suggest downstream applications to study the impact of substrate stiffness on DC function and metabolism. The protocol could be adapted to study other mechanosensitive immune cell subsets. For complete details on the use and execution of this protocol, please refer to Chakraborty et al. (2021).

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