High-reproducible flow cytometric endothelial progenitor cell determination in human peripheral blood as CD34+/CD144+/CD3- lymphocyte sub-population.
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'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.
Redondo
Santiago
S
Department of Pharmacology, School of Medicine, Universidad Complutense, Madrid, Spain.
Hristov
Mihail
M
Gordillo-Moscoso
Antonio A
AA
Ruiz
Emilio
E
Weber
Christian
C
Tejerina
Teresa
T
D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34 / Q000032:analysis*; D017252:CD3 Complex / Q000032:analysis*; D015820:Cadherins / Q000032:analysis*; D002448:Cell Adhesion; D002454:Cell Differentiation; D048430:Cell Shape; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D016131:Lymphocyte Subsets / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D015203:Reproducibility of Results; D013234:Stem Cells / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D000099152:Cadherin 5
D015703:Antigens, CD; D018952:Antigens, CD34; D017252:CD3 Complex; D015820:Cadherins; D000099152:Cadherin 5