Detection and quantification of mature circulating endothelial cells using flow cytometry and immunomagnetic beads: A methodological comparison

Patrick K. Y. Goon; Christopher J. Boos; Paul S. Stonelake; Andrew D. Blann; Gregory Y. H. Lip

doi:10.1160/TH06-04-0185

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2006; 96(01): 45-52
DOI: 10.1160/TH06-04-0185

Endothelium and Vascular Development

Schattauer GmbH

Detection and quantification of mature circulating endothelial cells using flow cytometry and immunomagnetic beads: A methodological comparison

Autoren

Patrick K. Y. Goon

¹Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, United Kingdom
Christopher J. Boos

¹Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, United Kingdom
Paul S. Stonelake

¹Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, United Kingdom
Andrew D. Blann

¹Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, United Kingdom
Gregory Y. H. Lip

¹Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, United Kingdom

Abstract

Summary

Mature circulating endothelial cells (CECs) are novel cellular markers of endothelial damage/dysfunction. The two main techniques of CEC enumeration are flow cytometry (FC) and immunomagnetic bead (IB) isolation. Both quantify CECs accurately, but a direct comparison of both methods has not been reported. We sought to assess the agreement between the two methods in two patient populations, and a group of healthy subjects, with emphasis given to methodological issues. We included 34 patients with acute coronary syndrome (ACS), 60 patients with primary breast cancer (PBC) and 30 healthy controls (HC). We quantified CECs using the IB method [CD146 and FITCUlex europaeus lectin-1] and FC [CD45, CD34 and CD146]. Bland-Altman plots suggested reasonable agreement (<5% of events >2 standard deviations from the mean) between FC and the IB methods for CEC quantification in whole blood in the two disease groups (ACS and PBC), but not among the HCs. There were no statistically significant differences in CEC levels by the two methods amongst all three patient groups. There is reasonable agreement between the FC and the IB methods for mature CEC quantification in whole blood, especially amongst disease groups. The agreement between the two methods appears to weaken in healthy controls, and at lower and higher absolute CEC counts.

Keywords

Circulating endothelial cells - endothelial cells - immunomagnetic beads - flow cytometry

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