Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2RS, Canada.
Division of Cellular and Molecular Biology, Diabetes Research Group, Toronto General Hospital Research Institute (TGHRI), University Health Network, Toronto, ON M5G 1L7, Canada.
STAR Protoc. 2022 Mar 17;3(2):101233. doi: 10.1016/j.xpro.2022.101233. eCollection 2022 Jun 17.
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).
聚二甲基硅氧烷水凝胶的机械性能可以进行调节，以模拟生理张力，这是免疫学研究中一个被低估的环境参数。我们描述了一种制备具有生物相关基质硬度的 PDMS 涂层组织培养板的工作流程，并使用这些水凝胶板来培养分离的原发性脾脏 CD11c+树突状细胞（DC）。最后，我们提出了下游应用，以研究基质硬度对 DC 功能和代谢的影响。该方案可以进行修改，以研究其他机械敏感免疫细胞亚群。如需了解本方案的使用和实施的详细信息，请参考 Chakraborty 等人（2021 年）的研究。
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