Enrichment of pluripotent hemopoietic progenitor cells from human bone marrow.

Enrichment of pluripotent hemopoietic progenitor cells from human bone marrow.
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富集人骨髓中的多能造血祖细胞。

DOI:
10.1182/blood.v64.4.774.774
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发表时间:
1984
期刊:
影响因子:
20.3
通讯作者:
M. Beard
M. Beard
中科院分区:
医学1区
文献类型:
--
作者:
M. Bodger;I. Hann;R. Maclean;M. Beard

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人骨髓多能造血祖细胞(CFU-GEMM,在甲基纤维素中形成混合造血祖细胞)通过使用单抗RFB-1在荧光激活细胞分选仪上进行阳性选择,扩增了90倍。根据细胞大小,用log90度光散射分离骨髓细胞,并用RFB-1抗原的相对荧光强度进一步分离含有CFU-GEMM的细胞部分。在RFB-1选择之前,通过耗尽骨髓中的T细胞和成熟的髓细胞,实现了进一步的浓缩,最高可达150倍。这些程序产生的细胞部分包含51%的原始细胞,2%的早幼粒细胞和47%的未分化(淋巴细胞样)单核细胞,尽管只有1%的细胞形成了混合集落。CFU-GEMM对RFB-1抗原呈强阳性,而形态可识别的红细胞、成髓细胞和早幼粒细胞呈弱RFB-1+。这表明,RFB-1抗原在骨髓细胞上的相对浓度与其成熟度呈负相关。用对数90度光散射分离骨髓后,CFU-GEMM的回收率大大提高,T细胞和成熟髓系细胞被去除,表明正常调节CFU-GEMM克隆效率的辅助细胞被去除。
Pluripotent hemopoietic progenitor cells (CFU-GEMM, cells forming mixed hemopoietic colonies in methylcellulose) from human bone marrow were enriched 90-fold by positive selection on the fluorescence-activated cell sorter using monoclonal antibody RFB-1. Bone marrow cells were separated by cell size, using log 90 degrees light scatter, and the cell fraction containing CFU-GEMM was further separated by relative fluorescence intensity for the RFB-1 antigen. Further enrichment, up to 150-fold, was achieved by depleting bone marrow of T cells and mature myeloid cells prior to RFB-1 selection. These procedures yield a cell fraction containing 51% blast cells, 2% promyelocytes, and 47% undifferentiated (lymphocyte-like) mononuclear cells, although only 1% of the cells formed a mixed colony. CFU-GEMM are strongly positive for the RFB-1 antigen, whereas morphologically identifiable erythroblasts, myeloblasts, and promyelocytes are weakly RFB-1+. This suggests that the relative concentration of the RFB-1 antigen on bone marrow cells is inversely related to their maturity. The greatly increased recovery of CFU-GEMM after the separation of bone marrow by log 90 degrees light scatter and the removal of T cells and mature myeloid cells suggested that accessory cells that normally regulate the cloning efficiency of CFU-GEMM were removed.
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