Related Experiment Video
Updated: Jun 24, 2025
10:25
Easy and Accurate Mechano-profiling on Micropost Arrays
Published on: November 17, 2015
11.1K
Multiplex, high-throughput method to study cancer and immune cell mechanotransduction
Abigail R Fabiano1, Spencer C Robbins1, Samantha V Knoblauch1
1Department of Biomedical Engineering, Vanderbilt University, 2414 Highland Ave, Nashville, TN, 37212, USA.
Communications Biology
|June 1, 2024
Summary
New high-throughput methods enable studying cellular responses to fluid shear stress (FSS) during cancer metastasis. These techniques enhance mechanotransduction research and improve workflow efficiency for analyzing cell activation and therapeutic effects.
Area of Science:
- Mechanobiology
- Cellular Mechanotransduction
- Cancer Metastasis Research
Background:
- Studying cellular mechanoresponses in cancer metastasis is challenging due to sample variability and complex existing techniques.
- Mechanotransduction, the process of cells converting physical stimuli like fluid shear stress (FSS) into biochemical signals, is crucial for metastasis.
- Current methods for applying FSS to cells are often low-throughput and labor-intensive.
Purpose of the Study:
- To develop high-throughput, semi-automated methods for applying controlled fluid shear stress (FSS) to cells.
- To establish protocols for live-cell staining and subsequent fixation, permeabilization, and intracellular analysis for flow cytometry.
- To demonstrate the utility of these methods in studying FSS-induced cellular responses in cancer and immunology models.
Main Methods:
- Utilized a VIAFLO96 multichannel pipetting device with custom-fitted 22G needles to achieve a 94-fold increase in maximum FSS compared to unmodified tips.
- Developed semi-automated protocols for live-cell staining, fixation, permeabilization, and intracellular processing for flow cytometry.
- Implemented a multiplex methodology for high-throughput analysis of cellular responses to FSS.
Main Results:
- Confirmed that FSS-induced Piezo1 activation enhances the pro-apoptotic effects of TRAIL therapeutics in prostate cancer cells.
- Demonstrated that FSS exposure (290 dyn cm⁻²) increases the activation of murine bone marrow-derived dendritic cells.
- Validated the high-throughput and multiplex capabilities of the developed mechanobiology workflow.
Conclusions:
- The presented high-throughput, semi-automated methods significantly improve the workflow for studying cellular mechanoresponses to FSS.
- These methodologies facilitate the investigation of mechanotransduction in cancer metastasis and immune cell activation.
- The developed approach offers a powerful tool for advancing mechanobiology research with enhanced efficiency and multiplexing capabilities.
