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用于研究细胞迁移的微加工柱阵列

[An array of microfabricated pillars to study cell migration].

作者信息

Buguin Axel, Chavrier Philippe, Ladoux Benoit, du Roure Olivia, Saez Alexandre, Silberzan Pascal

机构信息

Laboratoire de physico-chimie,UMR 168 CNRS, Institut Curie, 26 rue d'Ulm, 75005 Paris Cedex 05, France.

出版信息

Med Sci (Paris). 2005 Aug-Sep;21(8-9):765-7. doi: 10.1051/medsci/2005218-9765.

Abstract

Mechanical forces play an important role in various cellular functions, such as tumor metastasis, embryonic development or tissue formation. Cell migration involves dynamics of adhesive processes and cytoskeleton remodelling, leading to traction forces between the cells and their surrounding extracellular medium. To study these mechanical forces, a number of methods have been developed to calculate tractions at the interface between the cell and the substrate by tracking the displacements of beads or microfabricated markers embedded in continuous deformable gels. These studies have provided the first reliable estimation of the traction forces under individual migrating cells. We have developed a new force sensor made of a dense array of soft micron-size pillars microfabricated using microelectronics techniques. This approach uses elastomeric substrates that are micropatterned by using a combination of hard and soft lithography. Traction forces are determined in real time by analyzing the deflections of each micropillar with an optical microscope. Indeed, the deflection is directly proportional to the force in the linear regime of small deformations. Epithelial cells are cultured on our substrates coated with extracellular matrix protein. First, we have characterized temporal and spatial distributions of traction forces of a cellular assembly. Forces are found to depend on their relative position in the monolayer : the strongest deformations are always localized at the edge of the islands of cells in the active areas of cell protrusions. Consequently, these forces are quantified and correlated with the adhesion/scattering processes of the cells.

摘要

机械力在多种细胞功能中发挥着重要作用,如肿瘤转移、胚胎发育或组织形成。细胞迁移涉及黏附过程和细胞骨架重塑的动态变化,从而导致细胞与其周围细胞外介质之间产生牵引力。为了研究这些机械力,人们开发了许多方法,通过跟踪嵌入连续可变形凝胶中的珠子或微加工标记的位移来计算细胞与基质界面处的牵引力。这些研究首次对单个迁移细胞下的牵引力进行了可靠估计。我们开发了一种新型力传感器,它由使用微电子技术微加工的密集排列的软微米级柱子制成。这种方法使用通过硬光刻和软光刻相结合进行微图案化的弹性体基质。通过用光学显微镜分析每个微柱的偏转实时确定牵引力。实际上,在小变形的线性范围内,偏转与力成正比。上皮细胞在涂有细胞外基质蛋白的我们的基质上培养。首先,我们表征了细胞集合体牵引力的时间和空间分布。发现力取决于它们在单层中的相对位置:最强的变形总是位于细胞突起活跃区域中细胞岛的边缘。因此,这些力被量化并与细胞的黏附/散射过程相关联。

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