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1076029607304239v1
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First published on December 26, 2007, doi:10.1177/1076029607304239

Clinical and Applied Thrombosis/Hemostasis 2008;14:267.

A more recent version of this article appeared on July 1, 2008

Article

Further Insight Into the Heparin-Releasable and Glycosylphosphatidylinositol-Lipid—Anchored Forms of Tissue Factor Pathway Inhibitor

Paul E.R. Ellery, BSc, Kathy Hardy, PhD, Robert Oostryck, MSc, and Murray J. Adams, PhD*

University of Tasmania

* To whom correspondence should be addressed. E-mail: Murray.Adams{at}utas.edu.au.


  Abstract
The release of tissue factor pathway inhibitor (TFPI) from human umbilical vein endothelial cells (HUVECs) was investigated using heparin and phospholipase C. The experiment included incubating HUVECs with 0, 1, or 10 U/mL heparin diluted in Dulbecco Modified Eagle’s Medium plus 5% fetal calf serum for 1 or 24 hours. A statistically significant increase in TFPI activity levels was seen at 1 hour, but not at 24 hours. A 20-fold increase in the release of TFPI after phospholipase C treatment of HUVECs was demonstrated, confirming that it is glyco-sylphosphatidylinositol-lipid (GPI) anchored. Sequential treatment of HUVECs with phospholipase C and heparin was performed, and a trend was observed where GPI-anchored TFPI levels were increased after 1 hour of pretreatment with heparin but were decreased after 24 hours. Serum is a requirement for the heparin-dependent release of TFPI from HUVECs. Heparin pretreatment of HUVECs may affect levels of GPI anchored TFPI in a time and dose-dependent manner.


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