PDMS hydrogel-coated tissue culture plates for studying the impact of substrate stiffness on dendritic cell function

Megan Lee; Kevin Chu; Mainak Chakraborty; Nicholas Kotoulas; Masoud Akbari; Cynthia Goh; Xavier Clemente-Casares; Daniel A. Winer; Annie Shrestha; Sue Tsai

STAR Protocols (Jun 2022)

PDMS hydrogel-coated tissue culture plates for studying the impact of substrate stiffness on dendritic cell function

Megan Lee,
Kevin Chu,
Mainak Chakraborty,
Nicholas Kotoulas,
Masoud Akbari,
Cynthia Goh,
Xavier Clemente-Casares,
Daniel A. Winer,
Annie Shrestha,
Sue Tsai

Affiliations

Megan Lee: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada
Kevin Chu: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada
Mainak Chakraborty: Division of Cellular and Molecular Biology, Diabetes Research Group, Toronto General Hospital Research Institute (TGHRI), University Health Network, Toronto, ON M5G 1L7, Canada
Nicholas Kotoulas: Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada
Masoud Akbari: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada
Cynthia Goh: Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada
Xavier Clemente-Casares: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada
Daniel A. Winer: Division of Cellular and Molecular Biology, Diabetes Research Group, Toronto General Hospital Research Institute (TGHRI), University Health Network, Toronto, ON M5G 1L7, Canada; Buck Institute for Research on Aging, 8001 Redwood Boulevard, Novato, CA 94945, USA; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada
Annie Shrestha: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding author
Sue Tsai: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101233

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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