Comparison of two in vitro angiogenesis assays for evaluating the effects of netrin-1 on tube formation.

Netrin-1 is a neural guidance cue that also regulates vascular development. Controversial results, however, have been obtained concerning the roles of netrin-1 in vascular development both in vivo and in vitro. In the present study, two in vitro angiogenesis assays were compared to evaluate the effects of netrin-1 secreted by retrovirally transduced melanoma cells (Mel2a-netrin1) on tube formation. The results showed that there was no obvious difference in tube formation induced by conditioned media (CM) from the control, Mel2a-netrin1 and Mel2a cells in a matrigel assay. The results of another in vitro assay, in which endothelial cells were co-cultured with human fibroblasts, however, showed that Mel2a-netrin1 CM inhibited the tube formation, supposedly through blocking the elongation and coalescence of human umbilical vein endothelial cells (HUVECs). These results confirmed that the matrigel assay is not able to demonstrate the anti-angiogenic roles of netrin-1.