Table 1

Cells overexpressing VEGF have increased abilities to elicit PAE/KDR cell migration

MediumNo. of migrated cells after addition to the medium
NoneVEGF-neutralizing Ab Anti-mouse IgG2b.κ Ab
Medium alone138.2 ± 5.1 NDND
rhVEGF165638.7 ± 26.4 144.0 ± 4.9 628.2 ± 15.5
CM
 U87MG232.2 ± 5.3 218.6 ± 0.7 236.0 ± 7.0
 VEGF121561.0 ± 46.9 171.0 ± 6.5 542.7 ± 18.5
 VEGF165522.6 ± 27.7 193.1 ± 2.1 499.7 ± 24.5
 VEGF189
  Without heparin356.0 ± 8.2 NDND
  With heparin592.3 ± 22.8 NDND
  • In vitro migration of PAE/KDR cells in response to purified recombinant human VEGF165 (rhVEGF165, 10 ng/ml; R & D Systems) and to CM was measured in modified Boyden chambers. CM from the parental U87MG or their derivatives was diluted 1:100 with 0.1% BSA/Ham’s F-12 and tested in the absence or presence of either 1 μg/ml neutralizing anti-VEGF antibody or 1 μg/ml IgG2b.κ isotype control monoclonal antibody. Migrated cells were counted in 10 high-powered fields (×400 total magnification) per filter. All samples were examined in triplicate and data are shown as the mean numbers of migrated cells ± SEM. The assay was performed in triplicate for three independent sets of CM at least three times each with similar results. ND, not done.