High-reproducible flow cytometric endothelial progenitor cell determination in human peripheral blood as CD34 /CD144 /CD3- lymphocyte sub-population
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Although determination of circulating endothelial progenitor cell (EPC) in peripheral blood by flow cytometry is an emerging marker for cardiovascular medicine, a common standardized protocol is still not available, due to the low numbers achieved in peripheral blood. In the present paper we describe a novel technique for EPC quantification as CD34 /CD144 /CD3- cells within the lymphocyte gate, which increases the percentages of EPC positivity described before and also offers high intra-assay reproducibility. These improvements are based on a gating strategy for big-sized lymphocytes, smooth fixation and cytometric clearance of CD3 lymphocytes (T-cells). This last procedure is able to increase intra-assay Pearson%27s correlation from 0.8517 to 0.8908. Therefore, the technical setting described here offers a high-performance and clinically oriented EPC determination strategy in human peripheral blood. © 2008 Elsevier B.V. All rights reserved.
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Although determination of circulating endothelial progenitor cell (EPC) in peripheral blood by flow cytometry is an emerging marker for cardiovascular medicine, a common standardized protocol is still not available, due to the low numbers achieved in peripheral blood. In the present paper we describe a novel technique for EPC quantification as CD34%2b/CD144%2b/CD3- cells within the lymphocyte gate, which increases the percentages of EPC positivity described before and also offers high intra-assay reproducibility. These improvements are based on a gating strategy for big-sized lymphocytes, smooth fixation and cytometric clearance of CD3%2b lymphocytes (T-cells). This last procedure is able to increase intra-assay Pearson's correlation from 0.8517 to 0.8908. Therefore, the technical setting described here offers a high-performance and clinically oriented EPC determination strategy in human peripheral blood. © 2008 Elsevier B.V. All rights reserved.
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Endothelial progenitor cells; Flow cytometry; Markers CD3 antigen; CD34 antigen; vascular endothelial cadherin; adult; article; blood sampling; controlled study; female; flow cytometry; fluorescence activated cell sorting; human; human cell; lymphocyte subpopulation; male; peripheral blood stem cell; priority journal; Antigens, CD; Antigens, CD3; Antigens, CD34; Cadherins; Cell Adhesion; Cell Differentiation; Cell Shape; Cells, Cultured; Endothelial Cells; Female; Flow Cytometry; Humans; Immunophenotyping; Lymphocyte Subsets; Male; Middle Aged; Phenotype; Reproducibility of Results; Stem Cells; T-Lymphocytes
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