8 μm pore size transwells

Cell migration was determined using 8 μm pore size transwells (Corning Incorporated) for 24-well plates. One hundred thousand HepG2 cells expressing GFP/Fluc were seeded onto the upper insert of the Transwell chamber. MRC-5 fibroblasts were added at varying concentrations below as a chemottractant. After 24h at 37°C, non-invading cells were removed using a wet cotton swab. Migrating cells on the lower surface were imaged using fluorescence microscopy (×10) and were subsequently analyzed and quantified. Images were obtained at ×10 magnification.

For fluorescence-activated cell sorting, 1×10⁵ mast cells from thyroid tumor tissues were placed into the upper chambers (8-μm pore size transwells, Corning, USA). In the lower chambers, 50% TTCS or autologous 50% NTCS was added as the source of chemoattractants. Migration was calculated as the number of cells in the lower chamber after 24 h of culture at 37°C. In some cases, anti-SCF antibody (stem cell factor neutralizing antibody) or control IgG1 (20 μg/mL, IgG1) was added to the TTCS. Human recombinant cytokine SCF (100 ng/mL) and RPMI 1640 medium were added to the lower chambers as positive and blank controls.