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doi:10.22028/D291-33155
Title: | Micropatterned soft hydrogels to study the interplay of receptors and forces in T cell activation |
Author(s): | Zhang, Jingnan Zhao, Renping Li, Bin Farrukh, Aleeza Hoth, Markus Qu, Bin del Campo Bécares, Aránzazu |
Language: | English |
Title: | Acta biomaterialia |
Volume: | 119 |
Startpage: | 234 |
Endpage: | 246 |
Publisher/Platform: | Elsevier |
Year of Publication: | 2021 |
Publikation type: | Journal Article |
Abstract: | The analysis of T cell responses to mechanical properties of antigen presenting cells (APC) is experimentally challenging at T cell-APC interfaces. Soft hydrogels with adjustable mechanical properties and biofunctionalization are useful reductionist models to address this problem. Here, we report a methodology to fabricate micropatterned soft hydrogels with defined stiffness to form spatially confined T cell/hydrogel contact interfaces at micrometer scale. Using automatized microcontact printing we prepared arrays of anti-CD3 microdots on poly(acrylamide) hydrogels with Young's Modulus in the range of 2 to 50 kPa. We optimized the printing process to obtain anti-CD3 microdots with constant area (50 µm2, corresponding to 8 µm diameter) and comparable anti-CD3 density on hydrogels of different stiffness. The anti-CD3 arrays were recognized by T cells and restricted cell attachment to the printed areas. To test functionality of the hydrogel-T cell contact, we analyzed several key events downstream of T cell receptor (TCR) activation. Anti-CD3 arrays on hydrogels activated calcium influx, induced rearrangement of the actin cytoskeleton, and led to Zeta-chain-associated protein kinase 70 (ZAP70) phosphorylation. Interestingly, upon increase in the stiffness, ZAP70 phosphorylation was enhanced, whereas the rearrangements of F-actin (F-actin clearance) and phosphorylated ZAP70 (ZAP70/pY centralization) were unaffected. Our results show that micropatterned hydrogels allow tuning of stiffness and receptor presentation to analyze TCR mediated T cell activation as function of mechanical, biochemical, and geometrical parameters. |
DOI of the first publication: | 10.1016/j.actbio.2020.10.028 |
URL of the first publication: | http://www.sciencedirect.com/science/article/pii/S1742706120306176 |
Link to this record: | hdl:20.500.11880/30474 http://dx.doi.org/10.22028/D291-33155 |
ISSN: | 1878-7568 1742-7061 |
Date of registration: | 28-Jan-2021 |
Faculty: | M - Medizinische Fakultät NT - Naturwissenschaftlich- Technische Fakultät |
Department: | M - Biophysik NT - Chemie |
Professorship: | M - Prof. Dr. Markus Hoth NT - Prof. Dr. Aránzazu del Campo |
Collections: | SciDok - Der Wissenschaftsserver der Universität des Saarlandes |
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