Characterization of Extracellular Matrix (ECM) Produced by MC3T3 Cells Using Thickness Shear Mode (TSM) Resonators

Fang Li, Qing-Ming Wang, J.H.-C. Wang
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引用次数: 2

Abstract

Quartz thickness shear mode (TSM) resonators for monitoring the attachment and spreading of mammalian cells have been investigated in the past years. Recent studies have shown that the TSM resonator signal is not only contributed by cellular body closed to the resonator substrate, but also contributed by the extracellular matrix (ECM), which is a protein layer between the cellular body and the resonator surface. Therefore, the quartz TSM resonator is expected to provide a simple, quantitative and effective method to characterize the ECM produced by cells. In this study, we investigate the use of quartz TSM resonators to monitor the ECM production by cells in real time and to characterize the mechanical properties and the thickness of this protein layer. MC3T3-E1 fibroblasts are chosen in our study because they are an established cell line that is stable in culture from passage to passage and they can produce abundant collagenous matrix within a relatively short culture period. AT-cut quartz crystal resonators coated with gold electrodes on both sides are used in this study. An impedance analyzer is used for measuring the admittance spectra and total parallel capacitance of the resonators. Multilayer model is constructed to characterize the mechanical properties and thickness of the ECM layer by using a curve-fitting method. The results obtained by TSM resonator sensors show a good agreement with those obtained by chemical methods
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利用厚度剪切模式(TSM)谐振器表征MC3T3细胞产生的细胞外基质(ECM)
石英厚度剪切模式(TSM)谐振器用于监测哺乳动物细胞的附着和扩散在过去的几年里进行了研究。近年来的研究表明,TSM谐振器信号不仅由靠近谐振器底物的细胞体贡献,还由细胞体与谐振器表面之间的蛋白质层细胞外基质(extracellular matrix, ECM)贡献。因此,石英TSM谐振器有望提供一种简单、定量和有效的方法来表征细胞产生的ECM。在这项研究中,我们研究了使用石英TSM谐振器实时监测细胞产生的ECM,并表征该蛋白质层的力学特性和厚度。我们之所以选择MC3T3-E1成纤维细胞,是因为MC3T3-E1成纤维细胞是一种成熟的细胞系,在传代培养中稳定,在较短的培养时间内可以产生丰富的胶原基质。在本研究中使用了at切割的石英晶体谐振器,其两侧涂有金电极。用阻抗分析仪测量了谐振器的导纳谱和总并联电容。采用曲线拟合的方法,建立了多层模型来表征ECM层的力学性能和厚度。TSM谐振器传感器的测量结果与化学方法的测量结果吻合较好
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