Keese Charles R, Wegener Joachim, Walker Sarah R, Giaever Ivar
School of Science, Rensselaer Polytechnic Institute, 8th Street, Troy, NY 12180, USA.
Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1554-9. doi: 10.1073/pnas.0307588100. Epub 2004 Jan 27.
Confluent cell monolayers in tissue culture are fragile and can easily be mechanically disrupted, often leaving an area devoid of cells. This opening in the cell sheet is then repopulated, because the cells on the fringe of the damage, which are no longer contact-inhibited, move into the available space. This mechanical disruption is often done deliberately in a "wound-healing" assay as a means to assess the migration of the cells. In such assays, a scrape is made in the cell layer followed by microscopy to monitor the advance of the cells into the wound. We have found that these types of assays can also be accomplished electrically. In this approach, cells growing on small electrodes and monitored by using electric cell-substrate impedance sensing are subjected to currents, resulting in severe electroporation and subsequent cell death. After this invasive treatment, the electrode's impedance is again monitored to chart the migration and ultimate healing of the wound. We report here that this procedure to study cell behavior is both highly reproducible, quantitative, and provides data similar to that acquired with traditional measurements.
组织培养中汇合的细胞单层很脆弱，容易受到机械破坏，常常会留下一片无细胞区域。然后，细胞片层中的这个开口会重新被细胞占据，因为损伤边缘不再受接触抑制的细胞会迁移到这个可用空间中。这种机械破坏在“伤口愈合”实验中常常是故意进行的，以此作为评估细胞迁移的一种手段。在这类实验中，会在细胞层上刮出一道划痕，然后通过显微镜观察细胞向伤口处推进的情况。我们发现，这类实验也可以通过电的方式来完成。在这种方法中，生长在小电极上并通过细胞 - 基质阻抗传感进行监测的细胞会受到电流作用，导致严重的电穿孔及随后的细胞死亡。在这种侵入性处理之后，再次监测电极的阻抗，以记录伤口的迁移和最终愈合情况。我们在此报告，这种研究细胞行为的方法具有高度的可重复性、定量性，并且能提供与传统测量方法相似的数据。
