Martin H, Neumann M, Fache I, Fliedner T M, Pflieger H
Exp Hematol. 1985 Feb;13(2):79-86.
Two different density-gradient techniques were compared for separation of human blood-derived granulocyte-macrophage progenitor cells (CFUc) from immunocompetent cells using albumin or Percoll as gradient media. Mean CFUc enrichment by means of discontinuous albumin gradient was twofold, whereas Percoll gradients yielded about 20- to 40-fold enrichment of CFUc in relation to lymphocytes. The pH of gradient media proved to have a major influence on separation quality. Based on these results and the development of a new density-separation technique in a 600-ml plastic bag, replacing customary Ficoll-Hypaque gradients, a method for large-scale purification of CFUc from leukapheresis-derived leukocytes has been constructed, working in a closed, sterile system. About 11 X 10(9) mononuclear cells containing 6 X 10(5) CFUc could be processed within 10 h. Lymphocytes were reduced to less than 1% of their initial number, and an average of about 50% of the original CFUc could be recovered.
比较了两种不同的密度梯度技术，以使用白蛋白或 Percoll 作为梯度介质，从免疫活性细胞中分离人血来源的粒细胞 - 巨噬细胞祖细胞（CFUc）。通过不连续白蛋白梯度的平均 CFUc 富集倍数为两倍，而 Percoll 梯度使 CFUc 相对于淋巴细胞的富集倍数约为 20 至 40 倍。梯度介质的 pH 值被证明对分离质量有重大影响。基于这些结果以及在一个 600 毫升塑料袋中开发的一种新的密度分离技术（取代传统的 Ficoll - Hypaque 梯度），构建了一种从白细胞单采术获得的白细胞中大规模纯化 CFUc 的方法，该方法在封闭的无菌系统中运行。在 10 小时内可以处理约 11×10⁹个含有 6×10⁵个 CFUc 的单核细胞。淋巴细胞减少到其初始数量的不到 1%，并且平均可以回收约 50%的原始 CFUc。
