Swart K, Hagemeijer A, Löwenberg B
Exp Hematol. 1981 Jul;9(6):588-94.
Recently, PHA supplemented culture techniques have been introduced for growing colonies of myeloid leukemia cells. To prepare purified leukemic colony forming cell (CFC) suspensions for further studies, a discontinuous albumin density gradient separation method was applied to bone marrow and blood from patients with chronic myelocytic leukemia. It was found that the PHA-responding CFC were recovered, just as the leukocyte feeder layer stimulated CFC (Robinson CFC), from the light density fractions (1.056, 1.059 and 1.062 g/ml). Density profiles of the precursor cells forming colonies of Ph1 positive cells in the PHA-leukocyte feeder and Robinson assays appeared similar. T-lymphocyte progenitor cells, which also proliferate into colonies in the PHA-leukocyte feeder assay, were in majority harvested from the more dense fractions of the gradient. E-rosette tests and chromosome analysis were used to distinguish between leukemic and lymphocytic colonies. The density distributions of the PHA responsive leukemic CFC (Ph1 chromosome positive) and T-lymphocyte CFC (Ph1 negative) partially overlapped and a complete separation of leukemic and lymphocytic CFC was not achieved.
最近，已引入PHA补充培养技术来培养髓系白血病细胞集落。为了制备纯化的白血病集落形成细胞（CFC）悬液以进行进一步研究，对慢性粒细胞白血病患者的骨髓和血液应用了不连续白蛋白密度梯度分离法。结果发现，PHA反应性CFC与白细胞饲养层刺激的CFC（罗宾逊CFC）一样，从低密度组分（1.056、1.059和1.062 g/ml）中回收。在PHA-白细胞饲养层和罗宾逊试验中，形成Ph1阳性细胞集落的前体细胞的密度分布似乎相似。在PHA-白细胞饲养层试验中也增殖形成集落的T淋巴细胞祖细胞，大多从梯度中密度较高的组分中收获。采用E花环试验和染色体分析来区分白血病集落和淋巴细胞集落。PHA反应性白血病CFC（Ph1染色体阳性）和T淋巴细胞CFC（Ph1阴性）的密度分布部分重叠，未实现白血病CFC和淋巴细胞CFC的完全分离。
