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Localization of Tissue Factor Pathway Inhibitor In Endothelial Cells: Pitfalls and Challenges

Published online by Cambridge University Press:  02 July 2020

R. Olsen ,
J.-B. Hansen  and
P. Webster
R. Olsen
Affiliation:
Department of Electron Microscopy, Univeristy of Tromsø, N-9037 Tromsø, Norway
J.-B. Hansen
Affiliation:
Department of Medicine Faculty of Medicine, Univeristy of Tromsø, N-9037 Tromsø, Norway
P. Webster
Affiliation:
House Ear Institute, 2100 West Third Street, Los Angeles, CA90057
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Extract

Tissue Factor Pathway Inhibitor (TFPI) is a potent inhibitor of the extrinsic coagulation system, synthesized in endothelial cells. Membrane bound TFPI has recently been shown to play an important role in the degradation of activated coagulation factors in the endothelium. Comparison of subcellular TFPI localization patterns and TFPI production between Human Umbilical Vein Endothelial Cells (HUVEC) and an immortalized cell line (ECV 304) are presented here, as are the essential protocol modifications that we used to obtain labeling results by both light- and electron microscopy. We show that ECV 304 cells are similar to HUVEC in that TFPI is present in the Golgi complex (fig. 1) as well as in the endocytic pathway of ECV cells. Additionally, they secrete higher levels of TFPI into the culture medium than HUVEC. A colocalization of anti-TFPI with endocytosed BSA-gold, as well as with accumulated DAMP, demonstrated the presence of TFPI in the endocytic pathway.

Type
Blood/Immunology
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1124 - 1125
Copyright
Copyright © Microscopy Society of America

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