Ford Andrew J, Rajagopalan Padmavathy
ACS Biomater Sci Eng. 2018 Dec 10;4(12):3974-3982. doi: 10.1021/acsbiomaterials.8b01019. Epub 2018 Nov 9.
The cytoplasmic stiffness of cells plays a significant role during cell migration. As a cell migrates, differences in cytoplasmic properties occur that subsequently modulate migratory behavior. The properties of the substrate to which cells are adherent also play a role. To accurately measure the cytoplasmic stiffness of cells, we provide detailed instructions on how to assemble hydrogels that exhibit different elastic moduli, culturing cells on these substrates followed by a step-by-step process to measure and analyze the cytoplasmic properties of fibroblasts. In this study, we have measured the elastic moduli of cells at different locations to demonstrate how this property varies as a function of where the measurement is performed. The degree of anisotropy measured by the difference between cytoplasmic stiffness at the two edges of the cell also varied as a function of the elasticity of their underlying substrates. Larger differences in cytoplasmic stiffness between the leading and trailing edges were observed on substrates with a higher elastic modulus. The methods reported in this study can provide information on cellular properties, specifically, how the elastic modulus of cells can be probed and analyzed in vitro.
细胞的细胞质硬度在细胞迁移过程中起着重要作用。当细胞迁移时，细胞质特性会出现差异，进而调节迁移行为。细胞所附着的底物特性也发挥着作用。为了准确测量细胞的细胞质硬度，我们提供了关于如何组装具有不同弹性模量的水凝胶的详细说明，在这些底物上培养细胞，随后通过一个逐步的过程来测量和分析成纤维细胞的细胞质特性。在本研究中，我们测量了细胞在不同位置的弹性模量，以证明该特性如何随测量位置而变化。通过细胞两边细胞质硬度差异测量的各向异性程度也随其下方底物的弹性而变化。在具有较高弹性模量的底物上，前缘和后缘之间的细胞质硬度差异更大。本研究中报告的方法可以提供有关细胞特性的信息，具体而言，如何在体外探测和分析细胞的弹性模量。
