Inhibition of Glioma Cell Migration

Boyden chamber assay was performed using a 48-well chemotaxis chamber (Neuro-Probe, Gaithersburg, MD, USA) to confirm the inhibitory effect of CK on migration of C6 glioma cells. Scratch wound healing analyses were performed as previously reported [25 (link)]. Briefly, C6 glioma cells were suspended in serum-free medium at a density of 1 × 10⁶ cells/ml, and 50 µl of the cell suspension was loaded in the upper chamber wells. Treatment with SDF-1 with or without CK at different concentrations was performed in the lower chamber with incubation at 37℃ and 5% CO₂ atmosphere for 90 min. Cells on the lower surface of the membrane were fixed and stained using Diff Quick solutions. After staining, the migrated cells were photographed and counted under a microscope (Nikon, Tokyo, Japan). The migrated cells were measured using Image J software.