8 μm pore transwell migration chambers

Cell migration was evaluated by Transwell assay using the 8 μm-pore Transwell migration chambers (Corning, USA). Briefly, transfected cells were seeded in the upper portion of a chamber with serum-free medium. Complete medium filled in the lower chamber as an attractant. At 48 h after incubation, the non-migrated cells in the upper chamber were removed by cotton swabs. The migrated cells were stained with 0.1% crystal violet (Beyotime, Nantong, China) for 30 min, and then photographed and counted under optical microscope (Olympus, Tokyo, Japan).¹⁹ (link)

Cell proliferation assays were performed using a 5-ethynyl-2-deoxyuridine (EdU) cell proliferation assay kit (R11053.9, RiboBio, Guangzhou, China) according to the manufacturer’s instructions. Schwann cells were cultured in 96-well plates and spiked with 40 μg/ml of hAMSCs-exo or SCLCs-exo or an equal volume of PBS. After 6 h, the cells were observed under an inverted fluorescence microscope (Olympus, Tokyo, Japan).
Cell migration experiments were performed using 8-μm-pore Transwell migration chambers (Corning, NY, USA). A total of 2 × 10⁵ Schwann cells in 200 μl of serum-free medium were inoculated in the upper chamber of the Transwell plate. Subsequently, 40 μg/ml of hAMSCs-exo or SCLCs-exo or an equal volume of PBS was added to the upper chamber, while the lower chamber was filled with complete medium. The plates were incubated for 24 h before removing non-migrated cells from the upper chamber with a cotton swab. The migrated cells were stained with 0.1% crystalline violet (Beyotime, Nantong, China) for 30 min, photographed and counted under a light microscope (Olympus, Tokyo, Japan).