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[流式细胞术分析骨髓:形态学和免疫学方面]

[Analysis of bone marrow by flow cytometry: morphologic and immunologic aspects].

作者信息

Andreoni C, Rigal D, Bonnard M, Bernaud J

机构信息

Laboratoire d'Immunologie, Centre Régional de Transfusion Sanguine, Gerland, France.

出版信息

Pathol Biol (Paris). 1988 Dec;36(10):1183-91.

PMID:3070458
Abstract

Bone marrow aspirates from 28 healthy donors (18 adults, 10 children) were analysed by flow cytometry (FACS analyser) after purification of low density bone marrow cells (Ld BMC) on a density gradient (d = 1,077) and labeling with 23 anti-hematopoietic cells monoclonal antibodies. Based on physical properties the Ld BMC could be divided into four different populations called E, My, Mo et L including 13 +/- 8%, 33 +/- 15%, 12 +/- 5% and 42 +/- 14% of these cells respectively. The phenotypic analysis of these different populations allowed us to identify in E, erythrocytes (glycophorin A+, Rhesus D+ but negative for early erythroid differentiation markers like the transferrin receptor and/or the FA6-152 antigen); in My, the myeloid lineage (VIM2+, HLADR-); in Mo, the monocytic lineage (CD14+) and some myeloblasts (CD14-, VIM2+, HLADR+) and finally in L, an heterogeneous population including: 1) leucocyte cells, in which 27.3 +/- 9.0% are T cells labelled to the same extent by CD2, CD3, CD5 and CD6, 13.2 +/- 5.9% are B cells assessed by CD19 and CD20, 8.3 +/- 5.7% are Pré-B (CD10+), less than 5% are "natural killer" cells (CD16+ or Leu7+) and finally less than 6% are myelomonocytes (CD14+ or VIM12); 2) the erythroid lineage (Rhesus D+ = 43.7 +/- 12.9%, transferrin receptor and FA6-152+ 36.7 +/- 9.6%); 3) undifferentiated cells or progenitor cells (CD34+ = 6.5 +/- 3.5%); 4) cells unlabelled with any antibodies (approximatively 6%). We have not observed difference between adults and children bone marrow in regard to physical properties properties and all but also immunological markers. Indeed, a significant (p less than 0.02) higher proportion of B cells (CD19 and CD10) was observed in children. These data get from a large number of bone marrow could be used to quantify the imbalance of some bone marrow disorders.

摘要

对28名健康供体(18名成人,10名儿童)的骨髓穿刺样本进行分析,先在密度梯度(d = 1,077)上对低密度骨髓细胞(Ld BMC)进行纯化,然后用23种抗造血细胞单克隆抗体进行标记,之后通过流式细胞术(FACS分析仪)分析。基于物理特性,Ld BMC可分为四个不同群体,分别称为E、My、Mo和L,这些细胞分别占13±8%、33±15%、12±5%和42±14%。对这些不同群体的表型分析使我们能够在E群体中鉴定出红细胞(血型糖蛋白A+、恒河猴D+,但对早期红细胞分化标志物如转铁蛋白受体和/或FA6 - 152抗原呈阴性);在My群体中鉴定出髓系谱系(VIM2+、HLADR-);在Mo群体中鉴定出单核细胞谱系(CD14+)和一些成髓细胞(CD14-、VIM2+、HLADR+);最后在L群体中鉴定出一个异质群体,包括:1)白细胞细胞,其中27.3±9.0%是被CD2、CD3、CD5和CD6以相同程度标记的T细胞,13.2±5.9%是通过CD19和CD20评估的B细胞,8.3±5.7%是前B细胞(CD10+),少于5%是“自然杀伤”细胞(CD16+或Leu7+),最后少于6%是髓单核细胞(CD14+或VIM12);2)红细胞谱系(恒河猴D+ = 43.7±12.9%,转铁蛋白受体和FA6 - 152+ 36.7±9.6%);3)未分化细胞或祖细胞(CD34+ = 6.5±3.5%);4)未被任何抗体标记的细胞(约6%)。我们未观察到成人和儿童骨髓在物理特性以及免疫标志物方面存在差异。实际上,在儿童中观察到B细胞(CD19和CD10)的比例显著更高(p小于0.02)。这些来自大量骨髓的数据可用于量化某些骨髓疾病的失衡情况。

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