Molecular Human Reproduction, Vol. 7, No. 8, 771-777,
August 2001
© 2001 European Society of Human Reproduction and Embryology
Implantation and pregnancy |
In-vitro evidence of autocrine secretion of vascular endothelial growth factor by endothelial cells from human placental blood vessels
1 Division of Pharmacology and Chemotherapy and 2 Division of Pathology, Department of Oncology, Transplants and Advanced Technologies in Medicine, 3 Division of Gynaecology and Obstetrics, Department of Reproductive Medicine and Child Development, University of Pisa, Pisa, Italy
Abstract
Vascular endothelial growth factor (VEGF), a highly specific mitogen for vascular endothelial cells, is involved in placental vascular growth and remodelling. The aim of this study was to investigate whether placental endothelial cells secrete VEGF in an autocrine manner and if this secretion is correlated with endothelial cell growth. Blood vessels, excised from the apical surface of three human placentae, were sectioned into 40 fragments per placenta and cultured in fibrin gel matrix for 27 days. Immunohistochemical detection of placental endothelial cells was performed by positive staining with anti-human factor VIII-associated antigen and negative staining with anti-human 
-actin and desmin. To investigate the production and autocrine action of VEGF, VEGF concentrations in culture media were measured and the effect of an anti-VEGF neutralizing antibody on endothelial cell growth was observed. The results demonstrate that soluble VEGF is secreted by placental endothelial cells reaching a plateau from day 24 (68.74 ± 7.52 pg/ml) to day 27 (67.20 ± 6.28 pg/ml). Furthermore, VEGF concentrations in media collected on days 6, 12, 18, 21 and 27 of culture were found to be directly correlated to the sprouting parameter of endothelial cells, as calculated by image analysis on the same day (
P
< 0.001,
r
2
= 0.95
). The use of 10 and 100 ng/ml of a neutralizing antibody against human VEGF suppressed cell proliferation, compared to that observed in the untreated controls, by 74.8 ± 7.3 and 89.4 ± 3.9% respectively. In conclusion, this study reports the first evidence of autocrine secretion of VEGF by human placental endothelial cells and demonstrates the involvement of VEGF in endothelial cell growth within a fibrin gel culture.
endothelial cells/image analysis/immunohistochemistry/placental blood vessels/VEGF
Notes
4 To whom correspondence should be addressed at: Division of Pharmacology and Chemotherapy, Department of Oncology, Transplants and Advanced Technologies in Medicine, University of Pisa, Via Roma, 55, I-56126 Pisa, Italy. E-mail: m.deltacca{at}do.med.unipi.it
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