Yang Hsin-ya, La Thi Dinh, Isseroff R Rivkah
Department of Dermatology, Scool of Medicine, University of California, Davis;
Department of Dermatology, Scool of Medicine, University of California, Davis.
J Vis Exp. 2014 Dec 7(94):51973. doi: 10.3791/51973.
The physiological electric field serves specific biological functions, such as directing cell migration in embryo development, neuronal outgrowth and epithelial wound healing. Applying a direct current electric field to cultured cells in vitro induces directional cell migration, or galvanotaxis. The 2-dimensional galvanotaxis method we demonstrate here is modified with custom-made poly(vinyl chloride) (PVC) chambers, glass surface, platinum electrodes and the use of a motorized stage on which the cells are imaged. The PVC chambers and platinum electrodes exhibit low cytotoxicity and are affordable and re-useable. The glass surface and the motorized microscope stage improve quality of images and allow possible modifications to the glass surface and treatments to the cells. We filmed the galvanotaxis of two non-tumorigenic, SV40-immortalized prostate cell lines, pRNS-1-1 and PNT2. These two cell lines show similar migration speeds and both migrate toward the cathode, but they do show a different degree of directionality in galvanotaxis. The results obtained via this protocol suggest that the pRNS-1-1 and the PNT2 cell lines may have different intrinsic features that govern their directional migratory responses.
生理电场具有特定的生物学功能，如在胚胎发育中引导细胞迁移、神经元生长和上皮伤口愈合。在体外对培养的细胞施加直流电场可诱导细胞定向迁移，即趋电性。我们在此展示的二维趋电性方法是通过定制的聚氯乙烯（PVC）腔室、玻璃表面、铂电极以及使用配备有电动载物台的显微镜来实现的，细胞在该载物台上成像。PVC腔室和铂电极具有低细胞毒性，价格低廉且可重复使用。玻璃表面和电动显微镜载物台提高了图像质量，并允许对玻璃表面进行可能的修饰以及对细胞进行处理。我们拍摄了两种非致瘤性、SV40永生化前列腺细胞系pRNS - 1 - 1和PNT2的趋电性。这两种细胞系显示出相似的迁移速度，并且都向阴极迁移，但它们在趋电性中确实表现出不同程度的方向性。通过该方案获得的结果表明，pRNS - 1 - 1和PNT2细胞系可能具有不同的内在特征来支配它们的定向迁移反应。
