Moreover, this reversal was observed at the population level as all cells within the field of view switched direction within 30 minutes. The cells maintained rapid and directed migratory behaviour Abmole Neratinib toward the new cathode (Movie S5). Our results indicate that the cellular machinery required for NPC galvanotaxis can be actuated to induce migration – as well as reorganized to reverse migration – within 15 minutes of the dcEF onset or reversal, suggesting that de novo protein Abmole FK506 synthesis is not necessary for the process. We next asked if galvanotaxis is specific to undifferentiated NPCs or also a property of differentiated phenotypes. Neurospheres were plated into galvanotaxis chambers as described in the presence of 1% FBS for 69-72 hours to induce cell differentiation into mature neural phenotypes. Immunocytochemical analysis demonstrated that the majority of the cells expressed glial fibrillary acidic protein (GFAP) after 69 hours (Figure 3A), confirming that the NPCs had differentiated into astrocytes. This phenotype was maintained in FBS-cultured cells after 6 hours of dcEF exposure (Figure 3B). A rare subpopulation of cells continued to express nestin (Figure 3A, arrowhead) representing undifferentiated precursor cells. Differentiated cells were maintained in 1% FBS and either exposed or not exposed to a dcEF. Pre-differentiated cells exposed to a dcEF exhibited a mean 28.8365.08 mm displacement in the direction of the dcEF-vector at a mean velocity of 0.1460.02 mm/min (Figure 3C, 3D, S3, and Movie S6) Their directedness of migration was 20.2660.16, with a mean tortuosity value of 2.9260.25 (Figure 3E and 3F). This migratory behaviour did not differ significantly from that of differentiated cells that were not exposed to a dcEF or from undifferentiated cells in the absence of a dcEF (Figure 3C-3H, and Movie S7). Notably, post-dcEF labeling revealed that the differentiated cells (primarily astrocytes) aligned their processes perpendicular to the direction of the dcEF (Figure 3B).