Novel 3-D Macrophage Spheroid Model Reveals Reciprocal Regulation of Immunomechanical Stress and Mechano-Immunological Response
Year: 2024
Type: article
Abstract: Purpose: In many diseases, an overabundance of macrophages contributes to adverse outcomes. While numerous studies have compared macrophage phenotype after mechanical stimulation or with varying local stiffness, it is unclear if and how macrophages themselves contribute to mechanical forces in their microenvironment. Methods: Raw 264.7 murine macrophages were embedded in a confining agarose gel, where they proliferated to form spheroids over time. Gels were synthesized at various concentrations to tune the stiffness and treated with various growth supplements to promote macrophage polarization. The spheroids were then analyzed by immunofluorescent staining and qPCR for markers of proliferation, mechanosensory channels, and polarization. Finally, spheroid geometries were used to computationally model the strain generated in the agarose by macrophage spheroid growth. Results: Macrophages form spheroids and generate growth-induced mechanical forces (i.e., solid stress) within confining agarose gels, which can be maintained for at least 16 days in culture. Increasing agarose concentration restricts spheroid expansion, promotes discoid geometries, limits gel deformation, and induces an increase in iNOS expression. LPS stimulation increases spheroid growth, though this effect is reversed with the addition of IFN-γ. Ki67 expression decreases with increasing agarose concentration, in line with the growth measurements. Conclusions: Macrophages alone both respond to and generate solid stress. Understanding how macrophage generation of growth-induced solid stress responds to different environmental conditions will help to inform treatment strategies for the plethora of diseases that involve macrophage accumulation.
Authors: Alice Burchett, Saeed Siri, Jun Li, Xin Lu, Meenal Datta
Institution: University of Notre Dame
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,690
Funders: U.S. Department of Defense, Cancer Research Institute, Oak Ridge Associated Universities, Indiana Clinical and Translational Sciences Institute, University of Notre Dame, Mike and Josie Harper Cancer Research Institute, National Institutes of Health, Notre Dame Integrated Imaging Facility, University of Notre Dame, National Cancer Institute, National Institute of General Medical Sciences
Awards: G197919636, Converting Cold to Hot Tumor Microenvironment in Prostate Cancer by Targeting Chromatin Effector, G5641175177, TURN THE NEW ENEMY TO THE OLD FOE: NEUROENDOCRINE-TO-LUMINAL REDIFFERENTIATION, G729260546, DECODING AND DISRUPTING THE COUPLED CELLULAR PLASTICITY AND MYELOID CELL INSTIGATION IN METASTATIC PROSTATE CANCER, BENCH-BEDSIDE-BIDIRECTIONAL INVESTIGATION OF A NEW COMBINATORIAL IMMUNOTHERAPY STRATEGY TO TREAT PLATINUM-RESISTANT PENILE CANCER, TARGETING BASAL-LIKE PROSTATE CANCER WITH CADHERIN 3 ANTIBODY-DRUG CONJUGATE AS SINGLE AGENT AND IN COMBINATION WITH IMMUNOTHERAPY, G9000065865, G9000095542, G9000211313
OpenAlex ID: W4403395006