Bodger M P, Hann I M, Maclean R F, Beard M E
Blood. 1984 Oct;64(4):774-9.
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.
利用单克隆抗体RFB-1在荧光激活细胞分选仪上进行阳性选择，可将人骨髓中的多能造血祖细胞（CFU-GEMM，即在甲基纤维素中形成混合造血集落的细胞）富集90倍。通过对数90度光散射根据细胞大小分离骨髓细胞，然后根据RFB-1抗原的相对荧光强度进一步分离含有CFU-GEMM的细胞组分。在进行RFB-1选择之前，通过去除骨髓中的T细胞和成熟髓样细胞可实现进一步富集，高达150倍。这些操作得到的细胞组分含有51%的原始细胞、2%的早幼粒细胞和47%未分化的（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，而形态学上可识别的成红细胞、原粒细胞和早幼粒细胞RFB-1呈弱阳性。这表明骨髓细胞上RFB-1抗原的相对浓度与其成熟度呈负相关。通过对数90度光散射分离骨髓并去除T细胞和成熟髓样细胞后，CFU-GEMM的回收率大幅提高，这表明正常调节CFU-GEMM克隆效率的辅助细胞被去除了。
