Marley Stephen B, Lewis John L, Gordon Myrtle Y
Leukaemia Research Fund Centre for Adult Leukaemia, Department of Haematology, Imperial College Faculty of Medicine, London, UK.
Br J Haematol. 2003 May;121(4):643-8. doi: 10.1046/j.1365-2141.2003.04338.x.
Self-renewal is the most fundamental property of haemopoietic stem and progenitor cells. However, because of the need to produce differentiated cells, not all cell divisions involve self-renewal. We have used a colony replating assay to follow the fates of individual haemopoietic progenitor cell clones. For this, human myeloid colony-forming cells (CFCs) were cultured by standard methodology. Onset of proliferation and growth rates were established by a video recording method. Individual colonies were replated several times to document the rate of clonal extinction, and the numbers of secondary, tertiary and quaternary CFCs. The clonogenic population exhibited similar kinetics in terms of onset of proliferation and growth rate. Clonal extinction was progressive so that only 30 +/- 7% (mean +/- standard error of the mean; n = 4) of the original primary colonies formed quaternary colonies after the third replating step. However, individual primary CFCs that produced colonies throughout the experiment generated, on average, 40 +/- 8 secondary and tertiary CFCs overall. The values obtained in standard culture conditions were modified when granulocyte colony-stimulating factor (G-CSF) or G-CSF plus interleukin 3 were used to stimulate colony growth, showing that the kinetics of colony formation respond to extrinsic regulation. Examination of the replating potential of individual secondary colonies in the clones demonstrated that they generated different numbers of tertiary colonies. The data best fit a stochastic model of haemopoietic cell development where event probabilities can be modified by extracellular factors.
自我更新是造血干细胞和祖细胞最基本的特性。然而，由于需要产生分化细胞，并非所有细胞分裂都涉及自我更新。我们使用集落再接种试验来追踪单个造血祖细胞克隆的命运。为此，采用标准方法培养人髓系集落形成细胞（CFC）。通过录像方法确定增殖起始和生长速率。将单个集落多次再接种，以记录克隆灭绝率以及二级、三级和四级CFC的数量。克隆形成群体在增殖起始和生长速率方面表现出相似的动力学。克隆灭绝是渐进性的，因此在第三次再接种步骤后，只有30±7%（平均值±平均标准误差；n = 4）的原始一级集落形成了四级集落。然而，在整个实验过程中产生集落的单个一级CFC总体上平均产生40±8个二级和三级CFC。当使用粒细胞集落刺激因子（G-CSF）或G-CSF加白细胞介素3刺激集落生长时，在标准培养条件下获得的值会发生改变，这表明集落形成的动力学对外部调节有反应。对克隆中单个二级集落的再接种潜力进行检查表明，它们产生的三级集落数量不同。数据最符合造血细胞发育的随机模型，其中事件概率可由细胞外因子改变。
