Flow cytometric analysis of human bone marrow: I. Normal erythroid development.
Flow cytometric analysis of human bone marrow: I. Normal erythroid development.
复制标题
DOI：
10.1182/blood.v69.1.255.255
复制
发表时间：
1987
期刊：
影响因子：
20.3
通讯作者：
M. Loken;V. O. Shah;K. Dattilio;C. Civin
中科院分区：
文献类型：
--
作者：
M. Loken;V. O. Shah;K. Dattilio;C. Civin
应用流式细胞术结合物理特性、多种细胞表面抗原的表达和核酸含量来鉴定正常人骨髓中红系细胞的成熟差异。正常低密度骨髓细胞可分为四个群体，基于前向和直角光散射。在这四个骨髓亚群中的三个中发现了处于不同成熟阶段的红细胞。三种细胞表面标志物--HLe-1、转铁蛋白受体和血型糖蛋白--的顺序相关表达使我们能够研究从殖民地形成细胞到成熟红细胞的红系成熟。HLe-1在最早可识别的红系细胞上表达，并且随着细胞成熟而逐渐丢失。转铁蛋白受体在BFU-E期开始表达，网织红细胞晚期消失。转铁蛋白受体表达之前血型糖蛋白的表达，后者开始在形态学上可识别的红系前体CFU-E阶段后。与在成熟过程中逐渐丧失的HLe-1和转铁蛋白受体相反，一旦血型糖蛋白在细胞表面上最大程度地表达，它在成熟红细胞阶段保持恒定的量。虽然发展中的有核红系细胞在约normoblasts阶段的光散射特性类似的淋巴样细胞，这两种细胞类型可以解决细胞表面抗原的表达。正常成细胞血型糖蛋白阳性和HLe阴性，而淋巴样细胞表达HLe和Leu 4，Leu 11或Leu 12。细胞核酸含量的减少，首先对应于细胞核的挤出，其次对应于网状物的损失，其特征在于红系发育的后期阶段。这些特征和仪器可用于纯化不同发育阶段的红系细胞。
Flow cytometry was used to identify maturational differences of erythroid lineage cells in normal human bone marrow by combining physical characteristics, the expression of multiple cell surface antigens, and nucleic acid content. Normal low-density bone marrow cells could be divided into four populations, based on forward and right-angle light scattering. Erythroid cells, at different maturational stages, were found in three of these four marrow subpopulations. The sequentially correlated expression of three cell surface markers--HLe-1, transferrin receptor, and glycophorin--allowed us to study erythroid maturation from the colony forming cell to the mature erythrocyte. HLe-1 was expressed on the earliest identifiable erythroid cells and was progressively lost as the cells matured. Transferrin receptor began to be expressed at the BFU-E stage and disappeared at the late reticulocyte stage. Transferrin receptor expression preceded glycophorin expression, the latter beginning on morphologically recognizable erythroid precursors just after the CFU-E stage. In contrast to both HLe-1 and transferrin receptor, which were progressively lost during the maturational process, once glycophorin had been maximally expressed on the cell surface, it remained at constant quantities to the mature erythrocyte stage. Although developing nucleated erythroid cells at approximately the normoblast stage had light-scattering properties similar to those of lymphoid cells, these two cell types could be resolved by cell surface antigen expression. Normoblasts were glycophorin positive and HLe negative, whereas lymphoid cells expressed HLe and either Leu 4, Leu 11, or Leu 12. Decreases in cellular nucleic acid content, corresponding first to the extrusion of the nucleus and second to the loss of reticulum, characterized the later stages of erythroid development. These characteristics and instrumentation can be used to purify erythroid cells at various developmental stages.