Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry

Khan, Sameena S.; Solomon, Michael A.; McCoy, J. Philip

doi:10.1002/cyto.b.20040

Cited by 307 publications

(321 citation statements)

References 52 publications

(67 reference statements)

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“…Flow cytometry is a method of choice for quantification of ECs. Numerous attempts have been made to standardize flow cytometric protocols, but results from these studies show a high degree of variability among centers [8,27]. The reliable quantification of the ECs remains a technical challenge.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, the maturation process of ECs is not clearly understood, so the use of various marker combinations is necessary. The endothelial marker CD146, which is often used in EC analyses, is also expressed on active T lymphocytes [8,31]. The surface marker, CD144 (VE-cadherin), is expressed on both ECs and fetal hematopoietic stem cells [20,32,33].…”

Section: Discussionmentioning

confidence: 99%

“…The characterization and quantification of EC in the peripheral blood can provide a diagnostic or therapeutic advantage for diseases associated with endothelial damage because ECs are angiogenic and replace dysfunctional endothelium [4][5][6][7]. Under steady-state conditions in humans, the number of ECs in the circulation are relatively low (10 À4 to 10 À6 of all mononuclear cells present) [8][9][10]. Significant site-to-site variation has been observed with the current flow cytometry methods used to determine the percentage and absolute numbers of ECs.…”

Section: Introductionmentioning

confidence: 99%

“…The surface marker CD144 (vascular endothelial cadherin) labels ECs and fetal liver hematopoietic tissue cells [20]. These measurements show a wide variation in EC number because of rapid changes in endothelial surface markers with activation or maturation [8,[11][12][13]. In addition, we hypothesized that some ECs might be lost during the separation of mononuclear cells from other peripheral blood cells.…”

Section: Introductionmentioning

confidence: 99%

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Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

et al. 2007

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Several factors may influence the analysis of endothelial cells (ECs) by flow cytometry: separation of mononuclear cell, washing and centrifugation steps, panel of monoclonal antibodies, and the lack of standardization of gating technique. Therefore, the reliable quantification of ECs remains a technical challenge. The purpose of this study is to define a new flow cytometric protocol to characterize and quantitate ECs. In previous investigations, increased numbers of circulating ECs have been found in sickle cell disease. The patients with sickle cell disease might provide useful material for the study. We performed flow cytometry on whole blood from 20 normal controls and 31 patients with sickle cell disease (20 patients with steadystate disease and 11 patients with vaso-occlusive crises) using a lyse/no-wash procedure, specific and nonspecific antibody combinations (CD146, CD144, CD34, and CD117), and broad gating. This protocol produced much higher values for the number of circulating ECs (a mean of 2,396.55 ± 658.37 ECs/mL in controls vs 6,709.60 ± 1,772.32 ECs/mL in the steady-state group, or 18,213.50 ± 8,451 in the vaso-occlusive crises group, P < 0.001 for both), and also showed variable EC size and granularity, which may reflect activated, or early release ECs. This novel protocol performed comparably in terms of reproducibility, reliability, and dilution linearity with a previously described protocol. This approach has significant advantages for the characterization and quantitation ECs compatible with the pathophysiology. Using the specific antibodies, CD146 and CD144, together may give more informative EC data than the general approach used. Am. J. Hematol. 82:706-711, 2007. V

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

et al. 2007

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“…However, there have since been numerous publications describing correlations between CEC numbers and various human disease states, and CEC enumeration is gaining acceptance as a clinical biomarker of vascular disease [14]. Unfortunately, there is no consensus method for enumeration or immunophenotypic definition of human CEC/CEP [15]. In addition, very few publications exist on flow cytometry and CEC/CEP in animals.…”

Section: Endothelial Modelmentioning

confidence: 99%

Flow Cytometry in Preclinical Toxicology/Safety Assessment

McFarland¹,

Harkins

2010

Flow Cytometry in Drug Discovery and Development

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Identification of Endothelial Cells and Progenitor Cell Subsets in Human Peripheral Blood

et al. 2010

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An assay for circulating cell subsets in human peripheral blood by flow cytometry is used as a biomarker to determine cardiovascular disease risk and tumor responsiveness to chemotherapy since endothelial progenitor cells (EPCs) function in vasculogenesis and angiogenesis. Despite analytical advances in polychromatic flow cytometry (PFC), conventional approaches are routinely utilized to enumerate and isolate EPCs, which has led to varied results in clinical studies, potential cellular misidentification, and thus a lack of a plausible biological explanation for how purported EPCs function. Herein, a reproducible PFC protocol is provided to identify a rare circulating endothelial colony‐forming cell (ECFC) with proliferative potential, along with a population of circulating progenitor cells (CPCs) in which the ratio analysis distinguishes between healthy and disease populations. In sum, a reliable PFC protocol, which can be used to investigate the roles of human hematopoietic and endothelial elements in the growth and maintenance of the vasculature, is described. Curr. Protoc. Cytom. 52:9.33.1‐9.33.11. © 2010 by John Wiley & Sons, Inc.

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Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry

Cited by 307 publications

References 52 publications

Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

Flow Cytometry in Preclinical Toxicology/Safety Assessment

Identification of Endothelial Cells and Progenitor Cell Subsets in Human Peripheral Blood

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