Figure 5

The effect of antibody treatment on adhesion and transmigration of MC/car cells. (a) Binding of MC/car to RA-SNC77 cells. Biscarboxyethyl carboxyfluorescein-labeled MC/car cells (2 × 10⁵ cells/well) were plated onto a monolayer of RA-SNC77 cells (1 × 10⁴ cells/well) with or without mAb (20 g/ml) in a 96-well flat-bottomed culture plate. After 30 min of coculture, nonadherent cells were removed and the fluorescence intensity was measured. Results are expressed as the means ± standard deviation of three different experiments. ^*P < 0.05 compared with control, ^**P < 0.01 compared with control. (b) Transmigration of MC/car cells underneath RA-SNC77 cells. MC/car cells (1 × 10⁶ cells/well) were plated onto a monolayer of RA-SNC77 cells (1 × 10⁴ cells/well) with or without mAb (20 μg/ml) in a 12-well flat-bottomed culture plate. After 2 hours of coculture, MC/car cells bound to the surface of RA-SNC77 cells were removed, and the interaction between these cells was examined with a phase-contrast microscope. RA-SNC77 cells with more than three lymphoma cells beneath them were defined as positive. At least 200RA-SNC77 cells were counted in each experiment. Results are expressed as the means ± standard deviation of three different experiments. ^**P < 0.01 compared with control.