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- Title
Multiplex, high-throughput method to study cancer and immune cell mechanotransduction.
- Authors
Fabiano, Abigail R.; Robbins, Spencer C.; Knoblauch, Samantha V.; Rowland, Schyler J.; Dombroski, Jenna A.; King, Michael R.
- Abstract
Studying cellular mechanoresponses during cancer metastasis is limited by sample variation or complex protocols that current techniques require. Metastasis is governed by mechanotransduction, whereby cells translate external stimuli, such as circulatory fluid shear stress (FSS), into biochemical cues. We present high-throughput, semi-automated methods to expose cells to FSS using the VIAFLO96 multichannel pipetting device custom-fitted with 22 G needles, increasing the maximum FSS 94-fold from the unmodified tips. Specifically, we develop protocols to semi-automatically stain live samples and to fix, permeabilize, and intracellularly process cells for flow cytometry analysis. Our first model system confirmed that the pro-apoptotic effects of TRAIL therapeutics in prostate cancer cells can be enhanced via FSS-induced Piezo1 activation. Our second system implements this multiplex methodology to show that FSS exposure (290 dyn cm−2) increases activation of murine bone marrow-derived dendritic cells. These methodologies greatly improve the mechanobiology workflow, offering a high-throughput, multiplex approach. The authors present a multiplex, high-throughput method based on a commercially available multipipettor device to study mechanotransduction in cancer cells and immune cells in response to physiological fluid shear stress.
- Subjects
MECHANOTRANSDUCTION (Cytology); CANCER cells; MULTIPLEXING; SHEARING force; DENDRITIC cells
- Publication
Communications Biology, 2024, Vol 7, Issue 1, p1
- ISSN
2399-3642
- Publication type
Article
- DOI
10.1038/s42003-024-06327-x