Lu L, Bruno E, Briddell R A, Graham C D, Brandt J E, Hoffman R
Department of Medicine, Indiana Elks Cancer Research Center, Indiana University School of Medicine, Indianapolis 46223.
Behring Inst Mitt. 1988 Aug(83):181-7.
In order to study the effects of recombinant and purified hematopoietic growth factors on megakaryocyte (MK) progenitor cells (CFU-MK), enriched populations of human CFU-MK were isolated utilizing fluorescence activated cell sorting after labelling of cells with monoclonal antibodies exhibiting specificity to the My10 (HPCA-1) antigen and the major histocompatibility (MHC) class II (HLA-DR) locus. The CFU-MK cloning efficiency (CE) was 1.1 +/- 0.5% for cells expressing both high densities of My10 and low densities of HLA-DR (My10 DR+). This procedure resulted in an 18 fold increase in CE over NALT- cells. The effects of natural or recombinant human hematopoietic growth factors including erythropoietin (Epo), thrombocytopoiesis stimulating factor (TSF), interleukin 1 alpha (IL-1 alpha), granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (CSF-1), and interleukin 3 (IL-3) on MK colony formation by My10 DR+ cells were determined utilizing a defined medium assay system. Neither Epo, TSF, CSF-1, IL-1 alpha nor G-CSF alone augmented MK colony formation above baseline (2.5 +/- 0.8 per 5 x 10(3) My10 DR+ cells plated). By contrast, the addition of GM-CSF and IL-3 each increased CFU-MK colony formation with maximal stimulation occurring following the addition of 200 units/ml of IL-3 and 100 units/ml of GM-CSF. At maximal concentration, IL-3 had a greater ability to promote megakaryocyte colony formation than GM-CSF.
为了研究重组和纯化的造血生长因子对巨核细胞（MK）祖细胞（CFU-MK）的影响，在用对My10（HPCA-1）抗原和主要组织相容性（MHC）II类（HLA-DR）位点具有特异性的单克隆抗体标记细胞后，利用荧光激活细胞分选技术分离富集的人CFU-MK群体。对于同时表达高密度My10和低密度HLA-DR（My10 DR+）的细胞，CFU-MK克隆效率（CE）为1.1±0.5%。该方法使CE比NALT-细胞提高了18倍。利用限定培养基检测系统，测定了包括促红细胞生成素（Epo）、血小板生成刺激因子（TSF）、白细胞介素1α（IL-1α）、粒细胞集落刺激因子（G-CSF）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、巨噬细胞集落刺激因子（CSF-1）和白细胞介素3（IL-3）在内的天然或重组人造血生长因子对My10 DR+细胞形成MK集落的影响。单独使用Epo、TSF、CSF-1、IL-1α或G-CSF均未使MK集落形成超过基线水平（每接种5×10³个My10 DR+细胞，基线水平为2.5±0.8个）。相比之下，添加GM-CSF和IL-3均可增加CFU-MK集落形成，在添加200单位/ml的IL-3和100单位/ml的GM-CSF后刺激作用最大。在最大浓度时，IL-3促进巨核细胞集落形成的能力比GM-CSF更强。
