Chemotaxis Assay for Immune Cells

The induced mobility or chemotaxis of neutrophils, macrophages and lymphocytes was evaluated according to the method previously described [36 (link),70 (link)]. Cell suspensions were deposited in the upper compartment of a Boyden chamber separated by a filter of polycarbonate (3 μm in diameter; MercK, Madrid, Spain). The number of cells (neutrophils and lymphocytes) that migrated toward the chemoattractant agent, formyl-Met-Leu-Phe (fMLP, 10⁻⁸ M, Sigma-Aldrich, Madrid, Spain), deposited in the lower compartment of the chamber, was counted in the lower face of the filter that separates the two compartments of the chamber. This number is expressed as Chemotaxis Index (C.I.).