Human endothelium

Endovascular biopsy and molecular analysis

Lei Feng, David Stern, John Pile-Spellman

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)655-664
Number of pages10
JournalRadiology
Volume212
Issue number3
DOIs
StatePublished - Jan 1 1999

Fingerprint

Endothelium
Endothelial Cells
Biopsy
Iliac Artery
Polymerase Chain Reaction
Reverse Transcription
Blood Vessels
Lipopolysaccharides
Veins
Gene Expression
Thrombomodulin
Endovascular Procedures
E-Selectin
Vascular Cell Adhesion Molecule-1
Stainless Steel
Vascular Endothelium
von Willebrand Factor
Peptidyl-Dipeptidase A
Intercellular Adhesion Molecule-1
DNA-Directed RNA Polymerases

All Science Journal Classification (ASJC) codes

  • Radiology Nuclear Medicine and imaging

Cite this

Human endothelium : Endovascular biopsy and molecular analysis. / Feng, Lei; Stern, David; Pile-Spellman, John.

In: Radiology, Vol. 212, No. 3, 01.01.1999, p. 655-664.

Research output: Contribution to journalArticle

Feng, Lei ; Stern, David ; Pile-Spellman, John. / Human endothelium : Endovascular biopsy and molecular analysis. In: Radiology. 1999 ; Vol. 212, No. 3. pp. 655-664.
@article{a809f61907d547e79b3a0a37735bf063,
title = "Human endothelium: Endovascular biopsy and molecular analysis",
abstract = "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.",
author = "Lei Feng and David Stern and John Pile-Spellman",
year = "1999",
month = "1",
day = "1",
doi = "10.1148/radiology.212.3.r99au28655",
language = "English (US)",
volume = "212",
pages = "655--664",
journal = "Radiology",
issn = "0033-8419",
publisher = "Radiological Society of North America Inc.",
number = "3",

}

TY - JOUR

T1 - Human endothelium

T2 - Endovascular biopsy and molecular analysis

AU - Feng, Lei

AU - Stern, David

AU - Pile-Spellman, John

PY - 1999/1/1

Y1 - 1999/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0032873338&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032873338&partnerID=8YFLogxK

U2 - 10.1148/radiology.212.3.r99au28655

DO - 10.1148/radiology.212.3.r99au28655

M3 - Article

VL - 212

SP - 655

EP - 664

JO - Radiology

JF - Radiology

SN - 0033-8419

IS - 3

ER -