CD146‐based immunomagnetic enrichment followed by multiparameter flow cytometry: a new approach to counting circulating endothelial cells

Abstract

See also Shantsila E, Blann AD, Lip GYH. Circulating endothelial cells: from bench to clinical practice.This issue, pp 865-8.Summary. Background: Circulating endothelial cells (CECs) have emerged as non-invasive biomarkers of vascular dysfunction. The most widely used method for their detection is CD146-based immunomagnetic separation (IMS). Although this approach has provided consensus values in both normal and pathologic situations, it remains tedious and requires a trained operator. Objectives: Our objective was to evaluate a new hybrid assay for CEC measurement using a combination of preenrichment of CD146+ circulating cells and multiparametric flow cytometry measurement (FCM). Patients and methods: CECs were determined in peripheral blood from 20 healthy volunteers, 12 patients undergoing coronary angioplasty, and 30 renal transplant recipients, and blood spiked with cultured endothelial cells. CD146+ cells were isolated using CD146-coated magnetic nanoparticles and labeled using CD45-fluorescein isothiocyanate and CD146-PE or isotype control antibody and propidium iodide before FCM. The same samples were also processed using CD146-based immunomagnetic separation as the reference method. Results: The hybrid assay detected CECs, identified as CD45

Human Endothelial Progenitor Cells

“…Although numerous publications have proposed methods for resolution of circulating EPCs from circulating endothelial cells or other blood cell elements (Asahara et al 1997;Gehling et al 2000;Mancuso et al 2001Mancuso et al , 2009Bertolini et al 2003Bertolini et al , 2006Willett et al 2004Willett et al , 2005Abdul-Salam et al 2005;Khan et al 2005;Goon et al 2006;Duda et al 2007b;Van Craenenbroeck et al 2008;Widemann et al 2008), several recent publications have applied PFC to the identification of putative EPCs and have resolved the subsets into proangiogenic hematopoietic cells (Fig. 3) and circulating endothelial colony-forming cells (Estes et al 2010a,b;Schmidt-Lucke et al 2010).…”

Circulating endothelial cell count: a reliable marker of endothelial damage in patients undergoing hematopoietic stem cell transplantation

“…8,12,[16][17][18][19][20] However, due to their very low numbers and complex phenotype, several published techniques have shown different degrees of variability, reporting a wide range of CEC values (0-7900 CEC/mL) in healthy subjects. 12,[21][22][23][24] We prospectively correlated CEC count changes to GvHD onset and response to treatment in 90 allo-HSCT patients. Moreover, in this study, we used the commercially available rare cell isolation platform CellSearch system to identify and count CEC and potentially translate results into clinical practice.…”

Endothelial dysfunction and circulating endothelial cells in patients with chronic ischemic heart disease

“…Число ЦЭК во всех исследуемых группах было значительно выше общепринятой нормы, что, несомненно, подтверждает роль эндотели-альной дисфункции в развитии ИБС [12,13]. Достоверные различия числа ЦЭК между контрольной группой и группами пациентов с обструктивным поражением коронарного русла соотносятся с наличием атеросклероза.…”