A Multi-well Format Polyacrylamide-based Assay for Studying the Effect of Extracellular Matrix Stiffness on the Bacterial Infection of Adherent Cells

作者： Effie E Bastounis1, Fabian E Ortega1, Ricardo Serrano2, Julie A Theriot3 1Department of Biochemistry,Stanford University School of Medicine, 2Department of Mechanical and Aerospace Engineering,University of California San Diego, 3Departments of Biochemistry, Microbiology and Immunology and Howard Hughes Medical Institute,Stanford University School of Medicine

简介：

Overview

This study presents a polyacrylamide-based assay designed to investigate the impact of extracellular matrix stiffness on bacterial infection in adherent cells. The methodology allows for quantitative analysis of the biomechanical interactions between cells, their extracellular matrix, and pathogenic bacteria.

Key Study Components

Area of Science

• Biomechanics
• Cell Biology
• Microbiology

Background

• Understanding the role of mechanical forces in bacterial infection susceptibility is crucial.
• The emerging field of host-pathogen biomechanics explores these interactions.
• Current methods lack the ability to quantitatively assess these biomechanical factors.
• This study aims to fill that gap with a new assay.

Purpose of Study

• To characterize the effect of extracellular matrix stiffness on bacterial infection quantitatively.
• To investigate how mechanical forces influence the susceptibility of host cells to bacterial infection.
• To develop a methodology that allows for high-resolution imaging and automated screening.

Methods Used

• Preparation of polyacrylamide hydrogels with tunable stiffness.
• Seeding of human microvascular endothelial cells on the hydrogels.
• Infection of cells with Listeria monocytogenes and subsequent analysis.
• Flow cytometry to quantify bacterial infection rates.

Main Results

• The assay successfully quantifies the impact of matrix stiffness on bacterial infection.
• High-resolution imaging reveals dynamic interactions between cells and bacteria.
• Mechanical properties of the extracellular matrix significantly influence infection susceptibility.
• Automated procedures enhance the efficiency of the assay.

Conclusions

• This study provides a robust platform for investigating host-pathogen interactions.
• The findings highlight the importance of biomechanical factors in bacterial infections.
• The methodology can be applied to further research in host-pathogen biomechanics.

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