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Vascular and Interventional Radiology |
1 From the Departments of Radiology (L.F., J.P.S.) and Surgery (D.M.S.), Columbia University, 177 Ft Washington Ave, New York, NY 10032. Received June 17, 1998; revision requested August 14; revision received September 23; accepted February 12, 1999. Address reprint requests to L.F. (e-mail: lf66@columbia.edu).
PURPOSE: To develop a safe and reproducible method for harvesting viable vascular endothelium to analyze gene expression at sites of vascular lesions.
MATERIALS AND METHODS: Coaxial curved stainless-steel guide wires were used to obtain samples of endothelial cells from large arteries and veins in 29 patients undergoing routine endovascular procedures. Three immunocytochemical markers were used to identify cells as endothelial. Cellular viability was evaluated in terms of cell membrane integrity, energy-dependent uptake of acetylated low-density lipoprotein, and cellular response to lipopolysaccharide. Single-cell reverse transcription polymerase chain reaction (PCR) and immunocytochemistry were used to study endothelial gene expression.
RESULTS: Cells with endothelial morphology and immunoreactivity for von Willebrand factor, thrombomodulin, and angiotensin-converting enzyme were consistently obtained from iliac and carotid arteries and large veins (average yield [± standard error] from 26 iliac arteries, 262 endothelial cells ± 45, 20%–30% of which were viable). These cells displayed induction of E-selectin messenger RNA at PCR after exposure to lipopolysaccharide. Expression of vascular cell adhesion molecule 1 transcripts in endothelial cells increased with patient age (P < .01), whereas expression of intercellular adhesion molecule 1 did not.
CONCLUSION: Viable endothelium can be obtained during routine angiography. Immunocytochemical and reverse transcription PCR analyses of these cells allowed determination of transcripts and proteins expressed by endothelium at sites of vascular lesions. Such information could aid in understanding mechanisms of vascular diseases and in clinical decision making.
Index terms: Arteries, biopsy, 9*.126, 9*.922 • Arteries, endothelium, 9*.922 • Arteries, iliac, 984.1261 • Biopsies, technology • Carotid arteries, 90.1261 • Molecular analysis, 9*.922 • Venae cavae, interventional procedure, 946.1261 • Pulmonary arteries, 944.1261
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