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

机译:使用力谱仪测量细胞粘弹性:蛋白质-细胞质相互作用的影响

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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.
机译:细胞黏附和流变特性在通过内皮屏障的细胞迁移中起着非常重要的作用,尤其是在炎症(白细胞)或癌转移(癌细胞)的情况下。为了表征细胞的粘弹性,我们设计了一种力谱仪(AFM),可以拉伸细胞,从而测量其流变性。这种定制的力谱仪可实现两种不同的可视化效果,一种是横向的,另一种是从下方的。它可以研究细胞拉伸过程中涉及的流变学的影响。为了测试我们的系统表征此类粘弹性质的能力,分析了ICAM-1转染的CHO细胞。测试了两种形式的ICAM-1:可以与细胞骨架相互作用的野生型ICAM-1,以及缺乏细胞质结构域并且不能与细胞骨架结合的突变形式。在这些细胞上进行的拉伸实验显示出长丝的形成。使用以前的细丝伸长模型,我们可以确定单个细胞的粘弹性。正如预期的那样,在野生型和突变型之间发现了不同的粘弹性成分,这表明ICAM-1和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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