Scratch wound assay to quantify and analyze migration rate of canine olfactory ensheathing and Schwann cells. (A) Schematic drawing of the scratch wound assay. Confluent cell monolayer (left drawing) is scratched using a pipette tip (middle). The right drawing shows the closure of the wound. The gap width is photographed at t0 and t8 and measured using ImageJ (red arrows) (B-D) Representative images of OB-OECs (B), OM-OECs (C) and Schwann cells (C stained for p75NTR and DAPI are shown. All cells displayed either a spindle-shaped and process-bearing phenotype or a more flattened phenotype with only short processes emanating from the cell soma. (E-F) Representative images of the scratch at t0 and after 8 h migration are shown exemplarily for OB-OECs. (G) Migration rate differs after 8 h between cell types. OB-OECs showed significant higher migration rates than OM-OECs or Schwann cells whereas OM-OECs and Schwann cells showed no significant difference. Histograms present mean ± SEM of 7 (60 scratches, OB-OECs), 7 (48 scratches, OM-OECs) and 5 experiments (46 scratches, SCs). Statistical analysis was performed using an ANOVA and the Newman-Keuls post-hoc test (**p < 0.01, *p < 0.05). Cells are stained against p75NTR (red) and DAPI (blue). Scale bars are 50 μm (B-D) and 200 μm (E and F).
Roloff et al. BMC Neuroscience 2013 14:141 doi:10.1186/1471-2202-14-141