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

doi:10.1038/s41598-018-24234-0

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 MariniPasquale 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

Risultato della ricerca: Contributo su rivista › Articolo in rivista › peer review

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 (CV_L), calculated on the basis of Median Fluorescence Intensity values of the most stable antigens that allowed CEC identification and count (CV_L of CD34bright on CEC ∼ 30%; CV_L of CD45 on Lymphocytes ∼ 20%). By aggregating data acquired from all sites, CEC numbers in the healthy population were captured (median_female = 9.31 CEC/mL; median_male = 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.

Lingua originale	Inglese
Numero di articolo	5823
Rivista	Scientific Reports
Volume	8
Numero di pubblicazione	1
DOI	https://doi.org/10.1038/s41598-018-24234-0
Stato di pubblicazione	Pubblicato - 1 dic 2018
Pubblicato esternamente	Sì

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10.1038/s41598-018-24234-0

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Lanuti, P., Simeone, P., Rotta, G., Almici, C., Avvisati, G., Azzaro, R., Bologna, G., Budillon, A., Di Cerbo, M., Di Gennaro, E., Di Martino, M. L., Diodato, A., Doretto, P., Ercolino, E., Falda, A., Gregorj, C., Leone, A., Losa, F., Malara, N., ... Miscia, S. (2018). A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges. Scientific Reports, 8(1), Articolo 5823. https://doi.org/10.1038/s41598-018-24234-0

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title = "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, 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.",

author = "Paola Lanuti and Pasquale Simeone and Gianluca Rotta and Camillo Almici and Giuseppe Avvisati and Rosa Azzaro and Giuseppina Bologna and Alfredo Budillon and \{Di Cerbo\}, Melania and \{Di Gennaro\}, Elena and \{Di Martino\}, \{Maria Luisa\} and Annamaria Diodato and Paolo Doretto and Eva Ercolino and Alessandra Falda and Chiara Gregorj and Alessandra Leone and Francesca Losa and Natalia Malara and Mirella Marini and Pasquale Mastroroberto and Vincenzo Mollace and Michele Morelli and Emma Muggianu and Giuseppe Musolino and Arabella Neva and Laura Pierdomenico and Silvia Pinna and Giovanna Piovani and Roca, \{Maria Serena\} and Domenico Russo and Lorenza Scotti and Tirindelli, \{Maria Cristina\} and Valentina Trunzo and Roberta Venturella and Carlo Vitagliano and Fulvio Zullo and Marco Marchisio and Sebastiano Miscia",

note = "Publisher Copyright: {\textcopyright} 2018 The Author(s).",

year = "2018",

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doi = "10.1038/s41598-018-24234-0",

language = "English",

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Lanuti, P, Simeone, P, Rotta, G, Almici, C, Avvisati, G, Azzaro, R, Bologna, G, Budillon, A, Di Cerbo, M, Di Gennaro, E, Di Martino, ML, Diodato, A, Doretto, P, Ercolino, E, Falda, A, Gregorj, C, Leone, A, Losa, F, Malara, N, Marini, M, Mastroroberto, P, Mollace, V, Morelli, M, Muggianu, E, Musolino, G, Neva, A, Pierdomenico, L, Pinna, S, Piovani, G, Roca, MS, Russo, D, Scotti, L, Tirindelli, MC, Trunzo, V, Venturella, R, Vitagliano, C, Zullo, F, Marchisio, M & Miscia, S 2018, 'A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges', Scientific Reports, vol. 8, n. 1, 5823. https://doi.org/10.1038/s41598-018-24234-0

TY - JOUR

T1 - A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges

AU - Lanuti, Paola

AU - Simeone, Pasquale

AU - Rotta, Gianluca

AU - Almici, Camillo

AU - Avvisati, Giuseppe

AU - Azzaro, Rosa

AU - Bologna, Giuseppina

AU - Budillon, Alfredo

AU - Di Cerbo, Melania

AU - Di Gennaro, Elena

AU - Di Martino, Maria Luisa

AU - Diodato, Annamaria

AU - Doretto, Paolo

AU - Ercolino, Eva

AU - Falda, Alessandra

AU - Gregorj, Chiara

AU - Leone, Alessandra

AU - Losa, Francesca

AU - Malara, Natalia

AU - Marini, Mirella

AU - Mastroroberto, Pasquale

AU - Mollace, Vincenzo

AU - Morelli, Michele

AU - Muggianu, Emma

AU - Musolino, Giuseppe

AU - Neva, Arabella

AU - Pierdomenico, Laura

AU - Pinna, Silvia

AU - Piovani, Giovanna

AU - Roca, Maria Serena

AU - Russo, Domenico

AU - Scotti, Lorenza

AU - Tirindelli, Maria Cristina

AU - Trunzo, Valentina

AU - Venturella, Roberta

AU - Vitagliano, Carlo

AU - Zullo, Fulvio

AU - Marchisio, Marco

AU - Miscia, Sebastiano

PY - 2018/12/1

Y1 - 2018/12/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85045403019

U2 - 10.1038/s41598-018-24234-0

DO - 10.1038/s41598-018-24234-0

M3 - Article

SN - 2045-2322

VL - 8

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 5823

ER -

A standardized flow cytometry network study for the assessment of circulating endothelial cell physiological ranges

Abstract

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