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A novel flow cytometric approach to distinguish circulating endothelial cells from endothelial microparticles: Relevance for the evaluation of endothelial dysfunction

2012; Elsevier BV; Volume: 380; Issue: 1-2 Linguagem: Inglês

10.1016/j.jim.2012.03.007

1872-7905

Paola Lanuti, Francesca Santilli, Marco Marchisio, Laura Pierdomenico, Ester Vitacolonna, Eugenio Santavenere, A Iacone, Giovanni Davı̀, Mario Romano, Sebastianó Miscia,

Single-cell and spatial transcriptomics

Circulating endothelial cells (CEC) and endothelial microparticles (EMP) are emerging as markers of endothelial repair and activation/apoptosis. Although significant changes in the number of CEC and EMP in pathological conditions have been reported, their reliable identification and quantification still remain a technical challenge. Here, we present a novel methodology for the identification and quantitation of CEC and EMP based on multicolor flow cytometry. Using a lyse/no wash protocol, we observed that in 50 μl of peripheral blood, the large majority of events expressing an endothelial phenotype (CD45−/CD146+/CD34+) are due to non-nucleated particles (DRAQ5−) carrying mitochondrial activity (MitoTracker+) and, therefore, classified as EMP. We enumerated circulating EMP by single platform absolute count in a lyse/no wash four-color flow-cytometric procedure, which allowed the distinction, within the whole endothelial compartment, of EMP derived from endothelial progenitors (CD45−/CD146+/CD34+/CD117+) and from mature endothelial cells (CD45−/CD146+/CD34+/CD117−). A significant increase in both subsets was observed in patients with diabetes mellitus. Thus, this simple and highly reproducible method may be useful for monitoring endothelial dysfunction in clinical settings.