Swart K, Hagemeijer A, Löwenberg B
Leuk Res. 1982;6(1):55-62. doi: 10.1016/0145-2126(82)90043-1.
Investigation of leukemic colony-forming cells (CFC) in PHA-supplemented cultures requires removal of T lymphocyte precursors prior to culture. Using a method of discontinuous density gradient centrifugation with concurrent depletion of E-rosette forming cells, T lymphocytes were effectively separated from light density CML bone marrow and blood cell fractions. Consequently, in light density fractions (1.056 and 1.059 g/ml) pure leukemic colony growth was obtained in the PHA-leukocyte feeder (PHA-l.f.) assay. Fraction 1.062 g/ml also yielded pure leukemic colonies in most experiments. Comparison of the density distributions of leukemic PHA-l.f. CFC and Robinson CFC revealed that both CFC populations had congruent density profiles in most patients. In others PHA-l.f. CFC were found to be of somewhat higher density than Robinson CFC. The most striking divergence was apparent in a patient in blast crisis. The findings suggest that different subsets of precursor cells within the CML population proliferate in PHA-l.f. and Robinson colony methods. Both colony techniques are thus potentially useful for discriminating subpopulations of colony-forming cells in chronic myeloid leukemia.
在添加PHA的培养物中研究白血病集落形成细胞（CFC），需要在培养前去除T淋巴细胞前体。采用不连续密度梯度离心法并同时去除E花环形成细胞，可有效地将T淋巴细胞从低密度的慢性粒细胞白血病骨髓和血细胞组分中分离出来。因此，在低密度组分（1.056和1.059 g/ml）中，在PHA-白细胞饲养层（PHA-l.f.）试验中获得了纯白血病集落生长。在大多数实验中，1.062 g/ml的组分也产生了纯白血病集落。白血病PHA-l.f. CFC和罗宾逊CFC密度分布的比较显示，在大多数患者中，两个CFC群体具有一致的密度分布。在其他患者中，发现PHA-l.f. CFC的密度略高于罗宾逊CFC。最显著的差异在一例急变期患者中很明显。这些发现表明，慢性粒细胞白血病群体中的不同前体细胞亚群在PHA-l.f.和罗宾逊集落法中增殖。因此，这两种集落技术都可能有助于区分慢性粒细胞白血病中集落形成细胞的亚群。
