In angiogenesis associated with tissue repair and disease, fibrin and inflammatory mediators are often involved. We have used three-dimensional fibrin matrices to investigate the humoral requirements of human microvascular endothelial cells (hMVEC) to form capillary-like tubular structures. bFGF and VEGF165 were unable to induce tubular structures by themselves. Simultaneous addition of one or both of these factors with TNFalpha induced outgrowth of tubules, the effect being the strongest when bFGF, VEGF165, and TNFalpha were added simultaneously. Exogenously added u-PA, but not its nonproteolytic amino-terminal fragment, could replace TNFalpha, suggesting that TNFalpha-induced u-PA synthesis was involved. Soluble u-PA receptor (u-PAR) or antibodies that inhibited u-PA activity prevented the formation of tubular structures by 59-99%. epsilon-ACA and trasylol which inhibit the formation and activity of plasmin reduced the extent of tube formation by 71-95%. TNFalpha or u-PA did not induce tubular structures without additional growth factors. bFGF and VEGF165 enhanced of the u-PAR by 72 and 46%, but TNFalpha itself also increased u-PAR in hMVEC by 30%. Induction of mitogenesis was not the major contribution of bFGF and VEGF165 because the cell number did not change significantly in the presence of TNFalpha, and tyrphostin A47, which inhibited mitosis completely, reduced the formation of tubular structures only by 28-36%. These data show that induction of cell-bound u-PA activity by the cytokine TNFalpha is required in addition to the angiogenic factors VEGF165 and/or bFGF to induce in vitro formation of capillary-like structures by hMVEC in fibrin matrices. These data may provide insight in the mechanism of angiogenesis as occurs in pathological conditions.
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15 March 1996
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March 15 1996
Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity.
P Koolwijk,
P Koolwijk
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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M G van Erck,
M G van Erck
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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W J de Vree,
W J de Vree
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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M A Vermeer,
M A Vermeer
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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H A Weich,
H A Weich
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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R Hanemaaijer,
R Hanemaaijer
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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V W van Hinsbergh
V W van Hinsbergh
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
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P Koolwijk
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
M G van Erck
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
W J de Vree
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
M A Vermeer
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
H A Weich
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
R Hanemaaijer
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
V W van Hinsbergh
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1996) 132 (6): 1177–1188.
Citation
P Koolwijk, M G van Erck, W J de Vree, M A Vermeer, H A Weich, R Hanemaaijer, V W van Hinsbergh; Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity.. J Cell Biol 15 March 1996; 132 (6): 1177–1188. doi: https://doi.org/10.1083/jcb.132.6.1177
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