Four- and Five-Color Flow Cytometry Analysis of Leukocyte Differentiation Pathways in Normal Bone Marrow: A Reference Document Based on a Systematic Approach by the GTLLF and GEIL
Four- and Five-Color Flow Cytometry Analysis of Leukocyte Differentiation Pathways in Normal Bone Marrow: A Reference Document Based on a Systematic Approach by the GTLLF and GEIL
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DOI：
10.1002/cyto.b.20484
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发表时间：
2010-01-01
影响因子：
3.4
通讯作者：
Lacombe, Francis
中科院分区：
文献类型：
--
作者：
Arnoulet, Christine;Bene, Marie C.;Lacombe, Francis
背景资料：多参数流式细胞术（FCM）和日益复杂的分析软件的发展大大提高了血液系统疾病的探索。这些工具已广泛应用于白血病、淋巴瘤和骨髓增生异常，但方法非常异质。因此，没有广泛的参考文献报道正常人骨髓（BM）在多参数FCM中的特征。在这里，我们报告了一个参考分析程序，使用相关的抗体组合在正常人BM。方法：第一组23种抗体，文献综述后构建，进行了测试，在四色组合（包括CD45在每个）的30个BM样本。在评估数据后，将第二组22种抗体进一步应用于另外35个BM样品。对65份骨髓样本的所有列表模式进行了共同审查。建立了一个系统化的数据分析协议，包括三个主要的谱系和未分化celles.Results的双参数表示和颜色代码：这种策略已经允许获得一个参考图谱的分化抗原表达的相关模式在正常的人骨髓是在欧洲LeukemiaNet。这篇手稿介绍了如何构建这一图谱constructed.Conclusions：无论是战略和图谱可以证明是非常有用的正常参考，为确定白血病相关的免疫表型模式，分析骨髓增生异常，并最终调查的微小残留病的BM。(C)2009年临床细胞计数学会
Background: The development of multiparameter flow cytometry (FCM) and increasingly sophisticated analysis software has considerably improved the exploration of hematological disorders. These tools have been widely applied in leukaemias, lymphomas, and myelodysplasias, yet with very heterogeneous approaches. Consequently, there is no extensive reference document reporting on the characteristics of normal human bone marrow (BM) in multiparameter FCM. Here, we report a reference analysis procedure using relevant antibody combinations in normal human BM.Methods: A first panel of 23 antibodies, constructed after literature review, was tested in four-color combinations (including CD45 in each) on 30 samples of BM. After evaluation of the data, a second set of 22 antibodies was further applied to another 35 BM samples. All list-modes from the 65 bone marrow samples were reviewed collectively. A systematised protocol for data analysis was established including biparametric representations and color codes for the three major lineages and undifferentiated cells.Results: This strategy has allowed to obtain a reference atlas of relevant patterns of differentiation antigens expression in normal human BM that is available within the European LeukemiaNet. This manuscript describes how this atlas was constructed.Conclusions: Both the strategy and atlas could prove very useful as a reference of normality, for the determination of leukemia-associated immunophenotypic patterns, analysis of myelodysplasia and, ultimately, investigation of minimal residual disease in the BM. (C) 2009 Clinical Cytometry Society