Database-guided Flow-cytometry for Evaluation of Bone Marrow Myeloid Cell Maturation.

Autor: Aanei CM; Department of Hematology, University Hospital of Saint-Etienne; carmen.aanei@chu-st-etienne.fr., Jacob MC; Department of Immunology, University Hospital of Grenoble-Alpes., Veyrat-Masson R; Department Hematology, University Hospital of Clermont-Ferrand., Picot T; Department of Hematology, University Hospital of Saint-Etienne., Rosenthal-Allieri MA; Department of Immunology, University Hospital of Nice., Lhoumeau AC; Department of Biopathology, Institute Paoli-Calmettes., Ticchioni M; Department of Immunology, University Hospital of Nice., Dumezy F; Department of Hematology, University Hospital of Lille., Campos Catafal L; Department of Hematology, University Hospital of Saint-Etienne.
Jazyk: angličtina
Zdroj: Journal of visualized experiments : JoVE [J Vis Exp] 2018 Nov 03 (141). Date of Electronic Publication: 2018 Nov 03.
DOI: 10.3791/57867
Abstrakt: A working group initiated within the French Cytometry Association (AFC) was developed in order to harmonize the application of multiparameter flow cytometry (MFC) for myeloid disease diagnosis in France. The protocol presented here was agreed-upon and applied between September 2013 and November 2015 in six French diagnostic laboratories (University Hospitals of Saint-Etienne, Grenoble, Clermont-Ferrand, Nice, and Lille and Institut Paoli-Calmettes in Marseille) and allowed the standardization of bone marrow sample preparation and data acquisition. Three maturation databases were developed for neutrophil, monocytic, and erythroid lineages with bone marrow from "healthy" donor individuals (individuals without any evidence of a hematopoietic disease). A robust method of analysis for each myeloid lineage should be applicable for routine diagnostic use. New cases can be analyzed in the same manner and compared against the usual databases. Thus, quantitative and qualitative phenotypic abnormalities can be identified and those above 2SD compared with data of normal bone marrow samples should be considered indicative of pathology. The major limitation is the higher variability between the data achieved using the monoclonal antibodies obtained with the methods based on hybridoma technologies and currently used in clinical diagnosis. Setting criteria for technical validation of the data acquired may help improve the utility of MFC for MDS diagnostics. The establishment of these criteria requires analysis against a database. The reduction of investigator subjectivity in data analysis is an important advantage of this method.
Databáze: MEDLINE