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FICOLL DENSITY GRADIENT ISOLATION METHOD VS. DIRECT FLOW CYTOMETRIC QUANTIFICATION OF EPCS
Abstract:
Endothelial progenitor cells (EPCs) are bone-marrow derived cells involved in vascular homeostasis. In the present study, we assessed and compared two methods for EPCs isolation and quantification. The first method consisted of peripheral blood mononuclear cells (PBMC) isolation by Ficoll density gradient technique, followed by immunomagnetic labeling with specific antibodies beads (CD34, CD133 and VEGFR2/KDR) in order to enrich the target EPC subpopulation from the isolated PBMC. The selected immunolabelled cells were analyzed by flow cytometry. In the second method, whole blood was directly stained with specific antibodies and analyzed by flow cytometry. The aim of the study was to assess the efficiency and advantages of each method for EPCs quantification and also to evaluate the effect of immunomagnetic labeling with specific antibodies beads on EPCs quantification. The results have shown that there is not a significant difference between the EPCs number obtained by the two methods.

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