Fig. 2
Effect of mast cells on migratory capacity and proliferation of human lung fibroblasts, HFL-1. PBdMC enhanced the migratory capacity of HFL-1. Anti IgE stimulated PBdMC had a decreased pro-migratory effect compared to non-stimulated PBdMC (a). LAD2 cells, similarly to PBdMC, enhanced the migratory capacity of fibroblasts in a concentration-dependent manner, where the highest effect could be observed at 1 × 105 of LAD2 cells (b). There was no difference on the migration of HFL-1 in a co-culture with Anti IgE stimulated LAD2 compared to the non-stimulated LAD2 (c). The migratory capacity of HFL-1 cells was measured at 24, 48 and 72 h as the percentage of cell-occupied space compared to time 0 h when the scratch was made. LAD2 decreased proliferation of HFL-1 in co-cultures at 10% serum concentration, and did not have any effect on proliferation of HFL-1 at 0.4% serum concentration (d). The proliferation of HFL-1 was measured at 24, 48 72 h. The cell growth was determined using time-point 0 h as the reference point. (mean ± SD, n = 3 individual experiments with 4–6 technical replicates). Representative images of HFL-1 cells at 0 h (upper panels) and 72 h (lower panels). Images show 0.4% medium control, co-culture with PBdMC and co-culture with LAD2 (e) (mean ± SD, n = 3 individual experiments with 2 technical replicates in each experiment, *p < 0.05, **p < 0.01 and ***p < 0.001). The image of co-culture with PBdMC at time point 0 h, show recently added mast cells floating in the cell medium before attached to HFL-1 (e). Light microscopy images of co-cultures of HFL-1 (green arrow) and PBdMC (red arrow) (f), confocal microscopy images of HFL-1 and LAD2 co-cultures (g), Scanning electron microscopy (SEM) images HFL-1 and LAD2 (h)