Miller A M, Wu M C, Files N, Ingram M, Yunis A A
Stem Cells (1981). 1982;1(3):193-205.
We have attempted to purify human bone marrow granulocyte-macrophage progenitor cells utilizing purified stimulating factor, indirect immunofluorescence and cell sorting. These cells can be labelled by sequential incubation with colony-stimulating factor (CSF), rabbit anti-CSF antibody and fluorescently labelled goat anti-rabbit gamma globulin. Flow photometry demonstrated that binding of CSF to marrow cells varied with CSF concentration and duration of incubation. Marrow cells were pre-concentrated by incubation with carbonyl iron followed by Percoll gradient centrifugation and then labelled by this indirect immunofluorescence method. Sorting of these cells yielded up to 95-fold purification of colony-forming cells and fractions containing in excess of 10% colony- and cluster-forming cells.
我们尝试利用纯化的刺激因子、间接免疫荧光和细胞分选技术来纯化人骨髓粒细胞-巨噬细胞祖细胞。这些细胞可以通过依次与集落刺激因子（CSF）、兔抗CSF抗体以及荧光标记的山羊抗兔γ球蛋白孵育进行标记。流式光度法表明，CSF与骨髓细胞的结合随CSF浓度和孵育时间而变化。骨髓细胞先与羰基铁孵育进行预富集，然后通过Percoll梯度离心法处理，接着用这种间接免疫荧光法进行标记。对这些细胞进行分选可使集落形成细胞的纯化倍数高达95倍，且各组分中集落和集簇形成细胞的含量超过10%。
