Flow cytometric analysis of myelodysplasia: Pre-analytical and technical issues-Recommendations from the European LeukemiaNet.

Vincent H.j Van Der Velden ,Canan Alhan,Robin Ireland,Ulrika Johansson,Leonie Saft,Sharham Kordasti,Frank Preijers,Lisa Eidenshink‐Brodersen,Willemijn Hobo,Jeroen Te Marvelde,Uta Oelschlaegel,Nicolas Chapuis,Theresia M Westers,Arjan Van De Loosdrecht,Carolien Duetz,Peter Valent,Alan Dunlop,Anna Porwit,Marie C Béné,Peter Bettelheim,Patricia Font,Kate Burbury,Katherina Psarra,Dolores Subirá,Denise A Wells,Wolfgang Kern,Sergio Matarraz,Michael R Loken,Frauke Bellos,Eline M.p Cremers ,Orianne Wagner-Ballon ,Elisabeth H Weiß ,Michaëla Fontenay ,Matteo Giovanni Della Porta ,Alberto Órfão ,Kíyoyuki Ogata

doi:10.1002/cyto.b.22046

Abstract

Flow cytometry (FCM) aids the diagnosis and prognostic stratification of patients with suspected or confirmed myelodysplastic syndrome (MDS). Over the past few years, significant progress has been made in the FCM field concerning technical issues (including software and hardware) and pre-analytical procedures. Recommendations are made based on the data and expert discussions generated from 13 yearly meetings of the European LeukemiaNet international MDS Flow working group. We report here on the experiences and recommendations concerning (1) the optimal methods of sample processing and handling, (2) antibody panels and fluorochromes, and (3) current hardware technologies. These recommendations will support and facilitate the appropriate application of FCM assays in the diagnostic workup of MDS patients. Further standardization and harmonization will be required to integrate FCM in MDS diagnostic evaluations in daily practice.

Highlights

Flow cytometric immunophenotyping allows the identification, enumeration, and characterization of hematopoietic cells of distinct cell lineages and their differentiation stages in the bone marrow (BM) and peripheral blood (PB)
Samples processed at later time points may still be evaluable, but extra controls should be included to evaluate the quality of the sample (e.g., check scatter characteristics, percentage of dead cells, Flow cytometry (FCM) analysis of myelodysplastic syndrome (MDS) patients involves the granulocytic and monocytic lineages and analysis of the erythroid cell compartment, which provides additional valuable information (Cremers, et al, 2017; Mathis et al, 2013; Westers, et al, 2017)
FCM can contribute to the diagnosis and prognostication of MDS patients

Summary

| INTRODUCTION

Flow cytometric immunophenotyping allows the identification, enumeration, and characterization of hematopoietic cells of distinct cell lineages and their differentiation stages in the bone marrow (BM) and peripheral blood (PB). Samples processed at later time points may still be evaluable, but extra controls should be included to evaluate the quality of the sample (e.g., check scatter characteristics, percentage of dead cells, FCM analysis of MDS patients involves the granulocytic and monocytic lineages and analysis of the erythroid cell compartment, which provides additional valuable information (Cremers, et al, 2017; Mathis et al, 2013; Westers, et al, 2017). Antibodies that are informative of dysplastic features are summarized, and their diagnostic value is explained in detail in a separate paper in this issue of Clinical Cytometry These antibodies are combined in FCM panels that produce a maximum of information about differentiation of specific subsets for example CD34/CD117/HLA-DR, CD13/CD11b/CD16, and CD105/CD71/ CD36 for myeloid progenitor, neutrophilic and erythroid maturation, respectively. Recommended markers CD13, CD33, CD10, CD11b, CD15, CD38, CD7, CD56 HLA-DR, CD10, CD19 HLA-DR, CD13, CD33, CD11b, CD16, CD10, CD15, CD14, CD64, CD56 HLA-DR, CD13, CD33, CD11b, CD14, CD34, CD36, CD64, CD16, CD56, CD117 HLA-DR, CD36, CD71, CD105, CD13, CD33

HLADR CD45 TdT

Findings

10 | CONCLUSIONS