Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay

Erwin A. Kruger, Paul H. Duray, Maria G. Tsokos, David J. Venzon, Steven K. Libutti, Shannon C. Dixon, Michelle A. Rudek, James Pluda, Carmen Allegra, William D. Figg

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Endostatin has demonstrated potent antiangiogenic and antitumor activity in mouse models. We have investigated the ex vivo rat aortic ring assay and a human vein model to assess the biological activity of murine and human endostatin. Rat aortic rings were exposed to recombinant murine endostatin (Spodoptera frugipera; Calbiochem, San Diego, CA) or recombinant human endostatin (Pichia pastoris; EntreMed, Rockville, MD). After 5 days, murine endostatin (500 μg/ml) demonstrated inhibition of microvessel outgrowth with dose-dependent effects (down to 16 μg/ml). No significant inhibition was observed with human endostatin in the rat assay. Human endostatin at 250 and 500 μg/ml inhibited outgrowths from human saphenous vein rings after a 14-day incubation. Electron microscopy assessed the formation of basal lamina, confirming that the microvessels were progenitors of patent vessels. Immunostaining for Factor VIII or CD34 demonstrated that the microvessel cells were endothelial. BrdU incorporation assays supported the presence of proliferating endothelial cells, correlating with neovascularization from the aortic wall. We conclude that the rat aortic ring assay confirms the antiangiogenic activity of murine but not human endostatin, suggesting that the model may have species specificity. However, the human form shows biological activity against human vascular tissue. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)183-191
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume268
Issue number1
DOIs
StatePublished - Feb 5 2000
Externally publishedYes

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Endostatins
Microvessels
Rats
Assays
Endothelial cells
Bioactivity
Human Activities
Endothelial Cells
Species Specificity
Spodoptera
Pichia
Saphenous Vein
Factor VIII
Bromodeoxyuridine
Basement Membrane
Electron microscopy
Blood Vessels
Veins
Electron Microscopy
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Kruger, E. A., Duray, P. H., Tsokos, M. G., Venzon, D. J., Libutti, S. K., Dixon, S. C., ... Figg, W. D. (2000). Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay. Biochemical and Biophysical Research Communications, 268(1), 183-191. https://doi.org/10.1006/bbrc.1999.2018

Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay. / Kruger, Erwin A.; Duray, Paul H.; Tsokos, Maria G.; Venzon, David J.; Libutti, Steven K.; Dixon, Shannon C.; Rudek, Michelle A.; Pluda, James; Allegra, Carmen; Figg, William D.

In: Biochemical and Biophysical Research Communications, Vol. 268, No. 1, 05.02.2000, p. 183-191.

Research output: Contribution to journalArticle

Kruger, EA, Duray, PH, Tsokos, MG, Venzon, DJ, Libutti, SK, Dixon, SC, Rudek, MA, Pluda, J, Allegra, C & Figg, WD 2000, 'Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay', Biochemical and Biophysical Research Communications, vol. 268, no. 1, pp. 183-191. https://doi.org/10.1006/bbrc.1999.2018
Kruger, Erwin A. ; Duray, Paul H. ; Tsokos, Maria G. ; Venzon, David J. ; Libutti, Steven K. ; Dixon, Shannon C. ; Rudek, Michelle A. ; Pluda, James ; Allegra, Carmen ; Figg, William D. / Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay. In: Biochemical and Biophysical Research Communications. 2000 ; Vol. 268, No. 1. pp. 183-191.
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