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Investigating Cell-Substrate and Cell–Cell Interactions by Means of Single-Cell-Probe Force Spectroscopy

机译：通过单细胞探针力谱研究细胞底物和细胞间的相互作用

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摘要

Cell adhesion forces are typically a mixture of specific and nonspecific cell substrate and cell–cell interactions. In order to resolve these phenomena, Atomic Force Microscopy appears as a powerful device which can measure cell parameters by means of manipulation of single cells. This method, commonly known as cell-probe force spectroscopy, allows us to control the force applied, the area of interest, the approach/retracting speed, the force rate, and theudtime of interaction. Here, we developed a novel approach for in situ cantilever cell capturing and measurement of specific cell interactions. In particular, we present a new setup consisting of two different half-surfaces coated either with recrystallized SbpA bacterial cell surface layer proteinsud(S-layers) or integrin binding Fibronectin, on which MCF-7 breast cancer cells are incubated. The presence of a clear physical boundary between both surfaces benefits for a quick detection of the region under analysis. Thus, quantitative results about SbpA-cell and Fibronectin-cell adhesionudforces as a function of the contact time are described. Additionally, the importance of the cell spreading in cell–cell interactions has been studied for surfaces coated with two different Fibronectin concentrations: 20 lg/mL (FN20) and 100 lg/mL (FN100), which impact the number of substrate receptors.
机译：细胞粘附力通常是特异性和非特异性细胞底物与细胞间相互作用的混合物。为了解决这些现象，原子力显微镜似乎是一种功能强大的设备，可以通过操纵单个细胞来测量细胞参数。这种方法通常称为细胞探针力光谱法，它使我们能够控制所施加的力，感兴趣的区域，接近/收回速度，作用力速率以及相互作用的时间。在这里，我们开发了一种新方法，用于原位悬臂细胞捕获和特定细胞相互作用的测量。特别是，我们提出了一个新的设置，该设置由两个不同的半表面组成，这些表面覆盖有重结晶的SbpA细菌细胞表面层蛋白 ud（S-layers）或整合素结合的纤连蛋白，在其上孵育了MCF-7乳腺癌细胞。两个表面之间存在清晰的物理边界有利于快速检测分析区域。因此，描述了关于SbpA细胞和纤连蛋白细胞粘附力/力作为接触时间的函数的定量结果。此外，已经研究了涂有两种不同纤连蛋白浓度的表面在细胞之间相互作用中的重要性：20 lg / mL（FN20）和100 lg / mL（FN100），这会影响底物受体的数量。

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