Dynamic detection of natural killer cell-mediated cytotoxicity and cell adhesion by electrical impedance measurements
Dynamic detection of natural killer cell-mediated cytotoxicity and cell adhesion by electrical impedance measurements
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DOI：
10.1089/adt.2006.4.555
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发表时间：
2006-10-01
影响因子：
1.8
通讯作者：
Hansen, Anker Jon
中科院分区：
文献类型：
--
作者：
Glamann, Joakim;Hansen, Anker Jon
电阻抗测量是一种相对较新的实时、无标记的监测细胞粘附特性的方法。阻抗测量在底部装有金电极的组织培养孔中进行。活细胞的电极覆盖程度以及细胞膜和电极表面之间的结合强度决定了阻抗，在实时细胞电传感（RT-CES（TM），ACEA Biosciences，圣地亚哥，加利福尼亚州）中，阻抗被测量为细胞指数（CI）。我们发现，对于癌细胞，CI 与细胞数量呈线性相关，并且 CI 还与孔的涂层（层粘连蛋白-5）量相关。当自然杀伤 (NK) 细胞添加到贴壁癌细胞（靶细胞）中时，CI 迅速下降，具体取决于 NK 细胞：靶细胞的比例。根据 [H-3] 胸苷掺入测定，CI 的初始下降比靶细胞死亡更加明显。 CI 如此快速下降是由于发生凋亡的靶细胞的粘附和形态发生变化。细胞死亡程度和 CI 下降程度需要 6 个多小时才能达到可比性。我们还使用 A431 细胞和人表皮生长因子受体特异性抗体（Erbitux(R)，由德国达姆施塔特 Merck KGaA 制造）证明，RT-CES 可用于监测抗体依赖性细胞毒性。因此，RT-CES 是一种连续测定细胞数量和细胞粘附的便捷方法，并且可以提供 NK 细胞介导的细胞毒性作用的早期检测。
Measurement of electrical impedance is a relatively new real-time and label-free method for monitoring cell adhesive properties. Impedance measurements are performed in tissue culture wells in which the bottom is equipped with gold electrodes. The extent of electrode coverage by living cells as well as the strength of the bond between the cell membrane and the electrode surface determines the impedance, which in real-time cell electrical sensing (RT-CES (TM), ACEA Biosciences, San Diego, CA) is measured as the cell index (CI). We showed for carcinoma cells that CI was linearly correlated to the number of cells and that CI also was related to the amount of coating (laminin-5) of the wells. When natural killer (NK) cells were added to adherent carcinoma cells (target cells) CI declined rapidly dependent on the NK cell: target cell ratio. The initial decrease of CI was much more pronounced than target cell death as measured by [H-3] thymidine incorporation assay. Such a rapid fall of CI was due to changes in the adhesion and morphology of target cell undergoing apoptosis. It took more than 6 h before the extent of cell death and fall of CI were comparable. We also showed using A431 cells and an antibody specific for the human epidermal growth factor receptor (Erbitux((R)), manufactured by Merck KGaA, Darmstadt, Germany) that RT-CES could be used to monitor antibody-dependent cellular cytotoxicity. Thus RT-CES is a convenient way to continuously determine cell number and cell adhesion and may offer early detection of NK cell-mediated cytotoxic effects.