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Effect of cell density on the rate of the HUVEC morphogenesis

Because de novo c-met synthesis seems to be required for angiogenesis in vitro, the ECs that produce more c-met may undergo morphogenesis more readily than cells with a low c-met content. To test this hypothesis we analyzed the time course of angiogenesis in 3-dimensional collagen cultures obtained with confluent and sparse HUVECs. Indeed, as shown in Figure 6 the sparse HUVECs were characterized by the highest level of activity in morphogenesis. Just 1 hour after seeding in the 3-dimensional collagen matrix, virtually all of the sparse HUVECs had formed sprouts, whereas most of the confluent HUVECs were still round. After 3 hours, small sprouts could be seen in several confluent cells, whereas some short structures, formed by cells aligned end-to-end, were observed in the culture of sparse HUVECs. After 15 hours, the difference between these 2 types of culture was less great, probably due to the burst of c-met synthesis following the change of growth conditions.