Attachment of 5TGM1 myeloma cells to monolayer of mouse stromal cell line ST2 and effect of neutralizing antibodies to VCAM-1 and α4β1-integrin
. ST2 cells were cultured to confluency in 48-well culture plates.3H-Thymidine-labeled 5TGM1 cells (20 000 cells, 8654 ± 244 cpm) were seeded onto ST2 monolayer and incubated at 37°C for 1 hour in the absence or presence of 10 μg/mL of anti–VCAM-1 antibody (VCAM-1 Ab), anti-integrin α4β1-antibody (α4β1 Ab), both, or control IgG. After the incubation, nonadherent cells were removed by gentle washing with PBS, twice. Attached cells were lysed with 0.1 mol/L NaOH and radioactivity was determined by a liquid scintillation counter. Percentage of attached cells was calculated as radioactivity of adherent cells (cpm)/radioactivity of inoculated cells (cpm) × 100. Twenty μg/mL of these antibodies produced identical results (data not shown). Data are expressed as mean ± SE (n = 4). *Significantly different from IgG control (P < .01).
