Tucker D, Bol S, Kannourakis G
Department of Clinical Haematology & Oncology, Royal Children's Hospital, Melbourne, Australia.
Exp Hematol. 1993 Mar;21(3):469-74.
Hemopoietic cells that adhered to preformed selected marrow stromal cell layers were characterized on the basis of progenitor cell production capacity, colony-formation kinetics and forward light scatter (FLS) properties. It was shown that early hemopoietic cells attached to the stromal layers within 2 hours of incubation, and were responsible for the initial production of the more differentiated granulocyte-macrophage colony-forming cells (day 14 GM-CFC) in long-term cultures (LTC). In a clonogenic assay system, hemopoietic cells that adhere to stromal layers can be detected by the formation of small colonies of blast-like cells and are designated as stroma-adherent CFC. Cell fractionation on the basis of FLS and counting colonies on days 5, 14 and 21 revealed that there was a succession of colony formation, indicating that the stroma-adherent CFC consisted of subpopulations with different lag-phases before initiation of proliferation. Day 5, day 14 and day 21 stroma-adherent CFC were shown to have a high, intermediate and low FLS, respectively. The cells that produced GM-CFC by day 21 showed FLS properties similar to those of day 21 stroma-adherent CFC, suggesting a correlation between day 21 stroma-adherent CFC and CFC-producing cells in LTC. The CFC present on day 21 required the synergistic action of GM-CSF+IL-3 + stem cell factor (SCF) for optimal proliferation. The prolonged lag-phase, the low FLS and the multifactor-responsive progeny are properties similar to those reported for other early cells, and it is proposed that day 21 stroma-adherent CFC represent an early hemopoietic cell type whereas day 5 stroma-adherent CFC represent a more mature stage of differentiation.
根据祖细胞产生能力、集落形成动力学和前向光散射（FLS）特性，对黏附于预先形成的选定骨髓基质细胞层的造血细胞进行了表征。结果表明，早期造血细胞在孵育2小时内就附着于基质层，并在长期培养（LTC）中负责最初产生分化程度更高的粒细胞-巨噬细胞集落形成细胞（第14天的GM-CFC）。在克隆形成检测系统中，黏附于基质层的造血细胞可通过形成小的母细胞样细胞集落来检测，并被指定为基质黏附CFC。基于FLS进行细胞分级分离，并在第5天、第14天和第21天对集落进行计数，结果显示存在一系列集落形成，这表明基质黏附CFC由增殖开始前具有不同延迟期的亚群组成。第5天、第14天和第21天的基质黏附CFC分别显示出高、中、低FLS。到第21天产生GM-CFC的细胞显示出与第21天基质黏附CFC相似的FLS特性，这表明第21天基质黏附CFC与LTC中产生CFC的细胞之间存在相关性。第21天存在的CFC需要GM-CSF + IL-3 + 干细胞因子（SCF）的协同作用才能实现最佳增殖。延长的延迟期、低FLS和多因子反应性后代是与其他早期细胞报道的特性相似的特性，因此有人提出第21天的基质黏附CFC代表一种早期造血细胞类型，而第5天的基质黏附CFC代表一个更成熟的分化阶段。
