Immunosuppressant Effects on MSC Migration

Effect of immunosuppressants on MSC migration was examined in transwell plates with 8 μm pore filters (Corning, Cambridge, MA, USA) [14 (link)]. Upper wells were precoated with 0.1% gelatin (Sigma-Aldrich) for 2 h at 37°C. Upper wells were loaded with 2 × 10⁴ MSCs in 200 μl of medium and the lower wells with 500 μl of medium containing 10% fetal bovine serum and 100 ng/ml CXCL10 (R&D Systems). After 24 h, nonmigrated cells on the upper side of the filters were removed and the membranes were fixed in 10% formalin. MSCs that migrated to the lower side of the filter were stained with 0.5% crystal violet for 10 min and were counted under an inverted light microscope.