Scientific Reports (Apr 2018)
A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges
- 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
Affiliations
- Paola Lanuti
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Pasquale Simeone
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Gianluca Rotta
- BD Biosciences Italia
- Camillo Almici
- Laboratory for Stem Cells Manipulation and Cryopreservation, Department of Transfusion Medicine, Spedali Civili of Brescia
- Giuseppe Avvisati
- Hematology, Stem Cell Transplantation, Transfusion Medicine and Cellular Therapy, Department of Medicine, Campus Bio-Medico University Hospital
- Rosa Azzaro
- Transfusion Service, Department of Hematology-Oncology and Stem Cell Transplantation Unit
- Giuseppina Bologna
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Alfredo Budillon
- Experimental Pharmacology Unit, Department of Research, Istituto Nazionale Tumori- IRCCS G. Pascale
- Melania Di Cerbo
- Hematology, Stem Cell Transplantation, Transfusion Medicine and Cellular Therapy, Department of Medicine, Campus Bio-Medico University Hospital
- Elena Di Gennaro
- Experimental Pharmacology Unit, Department of Research, Istituto Nazionale Tumori- IRCCS G. Pascale
- Maria Luisa Di Martino
- Unit of Internal Medicine, Allergy and Clinical Immunology, Department of Medical Sciences “M. Aresu”, University of Cagliari Monserrato
- Annamaria Diodato
- Transfusion Service, Department of Hematology-Oncology and Stem Cell Transplantation Unit
- Paolo Doretto
- Clinical Pathology Laboratory, Department of Laboratory Medicine, AAS5, Pordenone Hospital
- Eva Ercolino
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Alessandra Falda
- Clinical Pathology Laboratory, Department of Laboratory Medicine, AAS5, Pordenone Hospital
- Chiara Gregorj
- Hematology, Stem Cell Transplantation, Transfusion Medicine and Cellular Therapy, Department of Medicine, Campus Bio-Medico University Hospital
- Alessandra Leone
- Experimental Pharmacology Unit, Department of Research, Istituto Nazionale Tumori- IRCCS G. Pascale
- Francesca Losa
- Unit of Internal Medicine, Allergy and Clinical Immunology, Department of Medical Sciences “M. Aresu”, University of Cagliari Monserrato
- Natalia Malara
- Department of Health Science University “Magna Graecia” of Catanzaro
- Mirella Marini
- Laboratory for Stem Cells Manipulation and Cryopreservation, Department of Transfusion Medicine, Spedali Civili of Brescia
- Pasquale Mastroroberto
- Department of Experimental and Clinical Medicine, University “Magna Graecia” of Catanzaro
- Vincenzo Mollace
- Department of Health Science University “Magna Graecia” of Catanzaro
- Michele Morelli
- Department of Obstetrics and Gynecology, University “Magna Graecia” of Catanzaro
- Emma Muggianu
- Unit of Internal Medicine, Allergy and Clinical Immunology, Department of Medical Sciences “M. Aresu”, University of Cagliari Monserrato
- Giuseppe Musolino
- Department of Experimental and Clinical Medicine, University “Magna Graecia” of Catanzaro
- Arabella Neva
- Laboratory for Stem Cells Manipulation and Cryopreservation, Department of Transfusion Medicine, Spedali Civili of Brescia
- Laura Pierdomenico
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Silvia Pinna
- Unit of Internal Medicine, Allergy and Clinical Immunology, Department of Medical Sciences “M. Aresu”, University of Cagliari Monserrato
- Giovanna Piovani
- Department Molecular Medicine and Translational, University of Brescia
- Maria Serena Roca
- Experimental Pharmacology Unit, Department of Research, Istituto Nazionale Tumori- IRCCS G. Pascale
- Domenico Russo
- Unit of Blood Diseases and Stem Cell Transplantation, University of Brescia
- Lorenza Scotti
- Department of Statistics and Quantitative Methods, University of Milano-Bicocca
- Maria Cristina Tirindelli
- Hematology, Stem Cell Transplantation, Transfusion Medicine and Cellular Therapy, Department of Medicine, Campus Bio-Medico University Hospital
- Valentina Trunzo
- Department of Health Science University “Magna Graecia” of Catanzaro
- Roberta Venturella
- Department of Obstetrics and Gynecology, University “Magna Graecia” of Catanzaro
- Carlo Vitagliano
- Experimental Pharmacology Unit, Department of Research, Istituto Nazionale Tumori- IRCCS G. Pascale
- Fulvio Zullo
- Department of Obstetrics and Gynecology, University “Magna Graecia” of Catanzaro
- Marco Marchisio
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- Sebastiano Miscia
- Department of Medicine and Aging Sciences, University “G.d’Annunzio”
- DOI
- https://doi.org/10.1038/s41598-018-24234-0
- Journal volume & issue
-
Vol. 8,
no. 1
pp. 1 – 10
Abstract
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.