Max Planck Institute for Dynamics and Self-Organization, Laboratory for Fluid Dynamics, Pattern Formation and Nanobiocomplexity (LFPN), Fassberg 17, D-37077 Goettingen, Germany.
Chem Res Toxicol. 2011 Sep 19;24(9):1494-506. doi: 10.1021/tx200115q. Epub 2011 Aug 22.
Cellular motility is the major driving force of numerous biological phenomena including wound healing, immune response, embryogenesis, cancer formation, and metastasis. We studied the response of epithelial FaDu monolayers cultured on gold electrodes of an acoustic resonator (quartz crystal microbalance, QCM) and impedance sensor (electric cell-substrate impedance sensing, ECIS) to externally applied chemical stimuli interfering with cytoskeleton organization. Epithelial cell motility of confluent monolayers is characterized by subtle cell shape changes and variations in the cell-substrate as well as cell-cell distance without net directionality of individual cells. The impact of small molecules such as cytochalasin D, phalloidin, and blebbistatin as well as paclitaxel, nocodazol, and colchicin on actin and microtubules organization was quantified by conventional sensors' readouts and by comparing the noise pattern of the signals which is attributed to cellular dynamics. The responsiveness of noninvasive and label-free techniques relying on cellular dynamics is compared to classical viability assays and changes of the overall impedance of ultrasmall electrodes or acoustic loads of a thickness shear mode resonator. Depending on the agent used, a distinct sensor response was found, which can be used as a fingerprint of the cellular response. Cytoskeletal rearrangements and nuclear integrity were corroborated by fluorescence microscopy and correlated to the readouts of QCM and ECIS.
细胞运动是许多生物学现象的主要驱动力，包括伤口愈合、免疫反应、胚胎发生、癌症形成和转移。我们研究了在声学谐振器（石英晶体微天平，QCM）和阻抗传感器（细胞-基底阻抗传感，ECIS）的金电极上培养的上皮 FaDu 单层对干扰细胞骨架组织的外部化学刺激的反应。在没有单个细胞的净方向性的情况下，细胞形态的细微变化以及细胞-基底和细胞-细胞距离的变化是融合单层上皮细胞运动的特征。细胞动力学归因于细胞动力学的信号噪声模式的比较，可以通过传统传感器的读数以及细胞松弛素 D、鬼笔环肽和 blebbistatin 以及紫杉醇、诺考达唑和秋水仙素等小分子对微丝和微管组织的影响来定量。比较了依赖于细胞动力学的非侵入性和无标记技术与经典活力测定以及超小型电极的整体阻抗变化或厚度剪切模式谐振器的声负载的响应。根据所用的试剂，发现了明显的传感器响应，可作为细胞反应的特征。细胞骨架重排和核完整性通过荧光显微镜得到证实，并与 QCM 和 ECIS 的读数相关。
