Abstract

The measurement of the forces at play in cell/cell adhesion uses a cell monolayer plated onto two-dimensional substrate like arrays of elastomeric microfabricated pillars. Unfortunately, the few attempts that have been done in this direction do not mimic the extracellular environment and lack knowledge on the correlation between the various types of cellular adhesions. Here, the authors suggest using a new experimental setup that more accurately simulates the three-dimensional (3D) environment of cells in tissues, using stretchable hexagonal monocellular 3D structures. The present article shows a way of fabricating these hexagonal biosensors, which are open structures made of biocompatible elastomeric polydimethylsiloxane (PDMS). The novelty of the whole fabrication process of these 3D PDMS structures consists in the use of a sacrificial silicon mold. An original mechanical PDMS planarization process is proposed. This article also describes a strategy for a selective functionalization of the 3D structure sidewalls where the cells must adhere.

Highlights

  • The organization of biological cells is sensitive to the biochemical composition of their environment and to its mechanical properties

  • Measuring the forces at play in cell/cell adhesion is challenging, since the use of a force sensor in a cell monolayer leads to the emergence of cell/extracellular matrix adhesions

  • The strategy we describe here uses a sacrificial silicon mold followed by an original planarization process that enables the opening of the PDMS structures

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Summary

Introduction

The organization of biological cells is sensitive to the biochemical composition of their environment and to its mechanical properties. The physical/ chemical nature of the external medium and its binding with the transmembrane proteins responsible for adhesion allow cells to probe the elastic properties of their environment.. The sensitivity of cells to the mechanical properties of the extracellular matrix arises from the mechanosensitive nature of cell adhesion.. Recent studies show that adhering cells actively probe the physical properties of the extracellular matrix by pulling onto it through its adhesive regions and respond by modulating their adhesion or their migration activity.. Measuring the forces at play in cell/cell adhesion is challenging, since the use of a force sensor in a cell monolayer leads to the emergence of cell/extracellular matrix adhesions. Isolated cells were plated onto 2D substrates with microfabricated pillars coated with cell/cell adhesion proteins or with pillars able to stress the cell as would do neighboring cells.. Isolated cells were plated onto 2D substrates with microfabricated pillars coated with cell/cell adhesion proteins or with pillars able to stress the cell as would do neighboring cells. cell geometry is known to influence gene expression and many cellular processes such as cell division or cell adhesion.

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