Zantl Roman, Horn Elias
ibidi GmbH, Martinsried, München, Germany.
Methods Mol Biol. 2011;769:191-203. doi: 10.1007/978-1-61779-207-6_13.
We present a microfabricated chamber designed for visualising and quantifying the chemotaxis of slow-migrating adherent mammalian cells such as cancer and endothelial cells. Most of the existing solutions for the investigation of chemotaxis are limited to fast migrating cells such as leukocytes or Dictyostelium discoideum. Here, we describe the details of an assay using the μ-Slide Chemotaxis to investigate the chemotactic response of human umbilical vein endothelial cells to a gradient of human vascular endothelial growth factor 165. In combination with phase contrast video microscopy and cell tracking, the trajectories of all single cells migrating in temporally stable gradients are derived. The resulting migration data are displayed and analysed in detail by several different parameters for quantifying chemotaxis. We found that with this tool the potential of chemoattractants to migration of mammalian cells as well as the impact of inhibitors to chemotaxis and migration can be evaluated.
我们展示了一种微制造的小室，其设计用于可视化和量化缓慢迁移的贴壁哺乳动物细胞（如癌细胞和内皮细胞）的趋化性。现有的大多数用于研究趋化性的方法都局限于快速迁移的细胞，如白细胞或盘基网柄菌。在此，我们描述了一种使用μ-Slide趋化性检测法来研究人脐静脉内皮细胞对人血管内皮生长因子165梯度的趋化反应的详细方法。结合相差视频显微镜和细胞追踪技术，可以得出所有在时间上稳定的梯度中迁移的单个细胞的轨迹。通过几个不同的用于量化趋化性的参数，对所得的迁移数据进行详细展示和分析。我们发现，使用这个工具可以评估趋化因子对哺乳动物细胞迁移的影响以及抑制剂对趋化性和迁移的影响。
