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使用力谱仪测量细胞粘弹性特性:蛋白质 - 细胞质相互作用的影响

Measuring cell viscoelastic properties using a force-spectrometer: influence of protein-cytoplasm interactions.

作者信息

Canetta Elisabetta, Duperray Alain, Leyrat Anne, Verdier Claude

机构信息

Université Grenoble I, Laboratoire de Spectrométrie Physique, Grenoble, France.

出版信息

Biorheology. 2005;42(5):321-33.

Abstract

Cell adhesive and rheological properties play a very important role in cell transmigration through the endothelial barrier, in particular in the case of inflammation (leukocytes) or cancer metastasis (cancer cells). In order to characterize cell viscoelastic properties, we have designed a force spectrometer (AFM) which can stretch cells thereby allowing measurement of their rheological properties. This custom-made force spectrometer allows two different visualizations, one lateral and one from below. It allows investigation of the effects of rheology involved during cell stretching. To test the ability of our system to characterize such viscoelastic properties, ICAM-1 transfected CHO cells were analyzed. Two forms of ICAM-1 were tested; wild type ICAM-1, which can interact with the cytoskeleton, and a mutant form which lacks the cytoplasmic domain, and is unable to associate with the cytoskeleton. Stretching experiments carried out on these cells show the formation of long filaments. Using a previous model of filament elongation, we could determine the viscoelastic properties of a single cell. As expected, different viscoelastic components were found between the wild type and the mutant, which reveal that the presence of interactions between ICAM-1 and the cytoskeleton increases the stiffness of the cell.

摘要

细胞黏附及流变学特性在细胞穿过内皮屏障的迁移过程中起着非常重要的作用,尤其是在炎症(白细胞)或癌症转移(癌细胞)的情况下。为了表征细胞的黏弹性特性,我们设计了一种力谱仪(原子力显微镜),它可以拉伸细胞,从而能够测量其流变学特性。这种定制的力谱仪允许两种不同的观察方式,一种是侧面观察,另一种是从下方观察。它能够研究细胞拉伸过程中涉及的流变学效应。为了测试我们的系统表征此类黏弹性特性的能力,对转染了细胞间黏附分子-1(ICAM-1)的中国仓鼠卵巢(CHO)细胞进行了分析。测试了两种形式的ICAM-1;一种是野生型ICAM-1,它可以与细胞骨架相互作用;另一种是突变形式,其缺乏细胞质结构域,无法与细胞骨架结合。对这些细胞进行的拉伸实验显示形成了长丝。使用先前的丝伸长模型,我们可以确定单个细胞的黏弹性特性。正如预期的那样,在野生型和突变体之间发现了不同的黏弹性成分,这表明ICAM-1与细胞骨架之间相互作用的存在增加了细胞的硬度。

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