Migration of HMC-1 cells to C3a (A) and C5a (B). Chemotactic response of HMC-1 cells to complement peptides was assayed in 48-well Boyden chambers. Various concentrations (1 nmol/L to 1 μmol/L) of C3a and C5a were placed in the lower wells of the chamber. The wells were covered with a laminin-coated (10 μg/mL) filter, and HMC-1 cells at 2 × 10 6 cells/mL were added to the upper wells of the chamber. Chambers were incubated for 150 minutes at 37°C, and filters were fixed, stained, and mounted using routine histologic methods. Mast cell migration was quantitated by counting the number of mast cells migrating through the filter. Spontaneous migration to medium/1% BSA served as control and was referred to as 100% migration. Data represent the mean ± SEM of 3 to 13 triplicate experiments for C3a (A; n = 3 to 13) and the mean ± SEM of 2 to 10 triplicate experiments for C5a (B; total n = 6 to 10).
