Scratch Assay for Cell Migration

Scratch assay was performed using silicone Ibidi inserts. Inserts were place in 12-well plate and 70 µL of standard BV2 medium with 35,000 cells were seeded in each compartment. After 24 h, inserts were removed, and the wells were filled with serum-free medium with or without LPS (100 ng/mL) [20 (link)] or ATP (300 µM) [71 (link)]. Cells were sham-treated or stimulated with ELF-EMS (20 min of 13.5 mT/60 Hz) 30 min after seeding. They were allowed to migrate for 24 h. Four pictures per sample were taken (20× magnification) 24 h after ELF-EMS with a Zeiss Model AxioVert 100 Inverted Microscope. The area occupied by migrated cells was quantified using Image J (version 1.53i) software [72 (link)]. Results are presented normalized to control culture.