Kontinuierliche Messung der zellgetriggerten Thrombinbildung – Entwicklung einer Methode zur Bestimmung der prokoagulatorischen Aktivität von Zellen

Under certain pathophysiological conditions endothelial cells expose procoagulant activity on their surface. A method for determination of the procoagulant activity of human umbilical vein endothelial cells was established. The cell-triggered generation of thrombin in plasma was measured by the use of the slow reacting substrate H-ß-Ala-Gly-Arg-pNA. The area under the thrombin generation curve serves as a measure for the procoagulant activity of the cells. The thrombin generation could be concentration-dependently inhibited by anti-tissue factor-antibodies and active site-blocked factor VIIa indicating a tissue factor mediated pathway. For endothelial cells there is a good correlation between the procoagulant activity measured by thrombin generation and the tissue factor antigen determined by an ELISA. The thrombin generation assay also was applied for the assessment of the procoagulant activity of other cell types such as smooth muscle cells, epithelial tumour cells or fibroblasts. The advantage of the presented method over usual methods is the assessment of the procoagulant activity of intact cells without great expenditure of work.