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一种能够在体外形成含原始细胞集落的人造血祖细胞的特性分析。

Characterization of a human hematopoietic progenitor cell capable of forming blast cell containing colonies in vitro.

作者信息

Brandt J, Baird N, Lu L, Srour E, Hoffman R

机构信息

Division of Hematology/Oncology, Indiana University School of Medicine, Indianapolis 46223.

出版信息

J Clin Invest. 1988 Sep;82(3):1017-27. doi: 10.1172/JCI113658.

Abstract

A hematopoietic cell (CFU-B1) capable of producing blast cell containing colonies in vitro was detected using a semisolid culture system. The CFU-B1 has the capacity for self-renewal and commitment to a number of hematopoietic lineages. Monoclonal antibody to the human progenitor cell antigen-1 (HPCA-1) and a monoclonal antibody against the major histocompatibility class II antigen (HLA-DR) were used with fluorescence activated cell sorting to phenotype the CFU-B1. The CFU-B1 was found to express My10 but not HLA-DR antigen; experiments using complement-dependent cytotoxicity to eliminate DR positive cells confirmed this finding. Pretreatment of marrow cells with two chemotherapeutic agents, 5-fluorouracil and 4-hydroperoxycyclophosphamide facilitated detection of CFU-B1 derived colonies, while diminishing or totally inhibiting colony formation by other hematopoietic progenitor cells. CFU-B1-derived colony formation was dependent upon the addition of exogenous hematopoietic growth factors. Media conditioned either by the human bladder carcinoma cell line 5637 or lectin stimulated leukocytes, as well as recombinant granulocyte-macrophage colony stimulating factor, interleukin 3 or interleukin 1 alpha promoted blast cell colony formation. By contrast, neither recombinant erythropoietin, recombinant interleukin 4, purified macrophage colony stimulating factor or recombinant granulocyte colony-stimulating factor alone promoted blast cell colony formation.

摘要

利用半固体培养系统检测到一种能够在体外产生含原始细胞集落的造血细胞(CFU - B1)。CFU - B1具有自我更新能力,并能分化为多种造血谱系。使用针对人祖细胞抗原 - 1(HPCA - 1)的单克隆抗体和针对主要组织相容性复合体II类抗原(HLA - DR)的单克隆抗体,通过荧光激活细胞分选对CFU - B1进行表型分析。发现CFU - B1表达My10但不表达HLA - DR抗原;使用补体依赖细胞毒性消除DR阳性细胞的实验证实了这一发现。用两种化疗药物5 - 氟尿嘧啶和4 - 氢过氧环磷酰胺预处理骨髓细胞,有助于检测CFU - B1衍生的集落,同时减少或完全抑制其他造血祖细胞的集落形成。CFU - B1衍生的集落形成依赖于外源性造血生长因子的添加。由人膀胱癌细胞系5637或凝集素刺激的白细胞条件培养液,以及重组粒细胞 - 巨噬细胞集落刺激因子、白细胞介素3或白细胞介素1α均能促进原始细胞集落形成。相比之下,单独的重组促红细胞生成素、重组白细胞介素4、纯化的巨噬细胞集落刺激因子或重组粒细胞集落刺激因子均不能促进原始细胞集落形成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d445/303616/ce353f7e14ee/jcinvest00081-0280-a.jpg

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