A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges
Abstract Circulating endothelial cells (CEC) represent a restricted peripheral blood (PB) cell subpopulation with high potential diagnostic value in many endothelium-involving diseases. However, whereas the interest in CEC studies has grown, the standardization level of their detection has not. Here...
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2018
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oai:doaj.org-article:4510c1edb29c4f32a18d18d2532c81152021-12-02T15:08:18ZA standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges10.1038/s41598-018-24234-02045-2322https://doaj.org/article/4510c1edb29c4f32a18d18d2532c81152018-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-24234-0https://doaj.org/toc/2045-2322Abstract Circulating endothelial cells (CEC) represent a restricted peripheral blood (PB) cell subpopulation with high potential diagnostic value in many endothelium-involving diseases. However, whereas the interest in CEC studies has grown, the standardization level of their detection has not. Here, we undertook the task to align CEC phenotypes and counts, by standardizing a novel flow cytometry approach, within a network of six laboratories. CEC were identified as alive/nucleated/CD45negative/CD34bright/CD146positive events and enumerated in 269 healthy PB samples. Standardization was demonstrated by the achievement of low inter-laboratory Coefficients of Variation (CVL), calculated on the basis of Median Fluorescence Intensity values of the most stable antigens that allowed CEC identification and count (CVL of CD34bright on CEC ~ 30%; CVL of CD45 on Lymphocytes ~ 20%). By aggregating data acquired from all sites, CEC numbers in the healthy population were captured (medianfemale = 9.31 CEC/mL; medianmale = 11.55 CEC/mL). CEC count biological variability and method specificity were finally assessed. Results, obtained on a large population of donors, demonstrate that the established procedure might be adopted as standardized method for CEC analysis in clinical and in research settings, providing a CEC physiological baseline range, useful as starting point for their clinical monitoring in endothelial dysfunctions.Paola LanutiPasquale SimeoneGianluca RottaCamillo AlmiciGiuseppe AvvisatiRosa AzzaroGiuseppina BolognaAlfredo BudillonMelania Di CerboElena Di GennaroMaria Luisa Di MartinoAnnamaria DiodatoPaolo DorettoEva ErcolinoAlessandra FaldaChiara GregorjAlessandra LeoneFrancesca LosaNatalia MalaraMirella MariniPasquale MastrorobertoVincenzo MollaceMichele MorelliEmma MuggianuGiuseppe MusolinoArabella NevaLaura PierdomenicoSilvia PinnaGiovanna PiovaniMaria Serena RocaDomenico RussoLorenza ScottiMaria Cristina TirindelliValentina TrunzoRoberta VenturellaCarlo VitaglianoFulvio ZulloMarco MarchisioSebastiano MisciaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-10 (2018) |
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Medicine R Science Q Paola Lanuti Pasquale Simeone Gianluca Rotta Camillo Almici Giuseppe Avvisati Rosa Azzaro Giuseppina Bologna Alfredo Budillon Melania Di Cerbo Elena Di Gennaro Maria Luisa Di Martino Annamaria Diodato Paolo Doretto Eva Ercolino Alessandra Falda Chiara Gregorj Alessandra Leone Francesca Losa Natalia Malara Mirella Marini Pasquale Mastroroberto Vincenzo Mollace Michele Morelli Emma Muggianu Giuseppe Musolino Arabella Neva Laura Pierdomenico Silvia Pinna Giovanna Piovani Maria Serena Roca Domenico Russo Lorenza Scotti Maria Cristina Tirindelli Valentina Trunzo Roberta Venturella Carlo Vitagliano Fulvio Zullo Marco Marchisio Sebastiano Miscia A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| description |
Abstract Circulating endothelial cells (CEC) represent a restricted peripheral blood (PB) cell subpopulation with high potential diagnostic value in many endothelium-involving diseases. However, whereas the interest in CEC studies has grown, the standardization level of their detection has not. Here, we undertook the task to align CEC phenotypes and counts, by standardizing a novel flow cytometry approach, within a network of six laboratories. CEC were identified as alive/nucleated/CD45negative/CD34bright/CD146positive events and enumerated in 269 healthy PB samples. Standardization was demonstrated by the achievement of low inter-laboratory Coefficients of Variation (CVL), calculated on the basis of Median Fluorescence Intensity values of the most stable antigens that allowed CEC identification and count (CVL of CD34bright on CEC ~ 30%; CVL of CD45 on Lymphocytes ~ 20%). By aggregating data acquired from all sites, CEC numbers in the healthy population were captured (medianfemale = 9.31 CEC/mL; medianmale = 11.55 CEC/mL). CEC count biological variability and method specificity were finally assessed. Results, obtained on a large population of donors, demonstrate that the established procedure might be adopted as standardized method for CEC analysis in clinical and in research settings, providing a CEC physiological baseline range, useful as starting point for their clinical monitoring in endothelial dysfunctions. |
| format |
article |
| author |
Paola Lanuti Pasquale Simeone Gianluca Rotta Camillo Almici Giuseppe Avvisati Rosa Azzaro Giuseppina Bologna Alfredo Budillon Melania Di Cerbo Elena Di Gennaro Maria Luisa Di Martino Annamaria Diodato Paolo Doretto Eva Ercolino Alessandra Falda Chiara Gregorj Alessandra Leone Francesca Losa Natalia Malara Mirella Marini Pasquale Mastroroberto Vincenzo Mollace Michele Morelli Emma Muggianu Giuseppe Musolino Arabella Neva Laura Pierdomenico Silvia Pinna Giovanna Piovani Maria Serena Roca Domenico Russo Lorenza Scotti Maria Cristina Tirindelli Valentina Trunzo Roberta Venturella Carlo Vitagliano Fulvio Zullo Marco Marchisio Sebastiano Miscia |
| author_facet |
Paola Lanuti Pasquale Simeone Gianluca Rotta Camillo Almici Giuseppe Avvisati Rosa Azzaro Giuseppina Bologna Alfredo Budillon Melania Di Cerbo Elena Di Gennaro Maria Luisa Di Martino Annamaria Diodato Paolo Doretto Eva Ercolino Alessandra Falda Chiara Gregorj Alessandra Leone Francesca Losa Natalia Malara Mirella Marini Pasquale Mastroroberto Vincenzo Mollace Michele Morelli Emma Muggianu Giuseppe Musolino Arabella Neva Laura Pierdomenico Silvia Pinna Giovanna Piovani Maria Serena Roca Domenico Russo Lorenza Scotti Maria Cristina Tirindelli Valentina Trunzo Roberta Venturella Carlo Vitagliano Fulvio Zullo Marco Marchisio Sebastiano Miscia |
| author_sort |
Paola Lanuti |
| title |
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| title_short |
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| title_full |
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| title_fullStr |
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| title_full_unstemmed |
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| title_sort |
standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges |
| publisher |
Nature Portfolio |
| publishDate |
2018 |
| url |
https://doaj.org/article/4510c1edb29c4f32a18d18d2532c8115 |
| work_keys_str_mv |
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