P38. Strong inhibitory effect of preeclampsia serum on angiogenesis using in vitro angiogenesis test

Abstract

Introduction The angiogenic response of preeclampsia patient serum samples were studied by using human in vitro angiogenesis test. The hypothesis was that preeclampsia patients’ serum may contain factors that disturb angiogenesis and that could explain formation of some of the adverse effects in patients. Objective To measure in vitro angiogenetic effects of serum samples from preeclamptic mothers and umbilical blood. Methods Serum samples were collected from ten primiparous preeclampsia patients and ten matched controls at gestational age of 28–38 weeks. After birth giving, also umbilical blood samples were taken. The test method consisted of a co-culture of human fibroblasts and endothelial cells in the culture medium with specific exogenous growth factors to induce formation of tubular structures and tubular networks. During the test, the co-culture was exposed to patient serum samples (dilution 1:15) and cultured for further 6 days (with one replenishment of the growth medium). After the exposure cytotoxicity was evaluated using mitochondrial viability (WST-1) assay. Tubular structures were evaluated microscopically after immunostaining with anti von Willebrand factor. Results There was no cytotoxicity shown with any of the samples, but the umbilical blood samples seemed to induce cell growth to some extent. There were no stimulatory effects found on tubule formation with any of the samples, but there was a strong inhibitory effect on tubule formation with serum from preeclampsia patients. Also umbilical blood from preeclamptic pregnancies showed inhibitory effect on tubule formation, but there was no clear correlation of serum from mothers compared to corresponding umbilical serum. No significant correlations to the severity of preeclampsia or any clinical features of the disease were noticed. Conclusion Both maternal and umbilical blood serum after established preeclampsia show inhibitory effects on angiogenesis, as tested by in vitro -model.