Terminal differentiation of murine erythroleukemia cells: physical stabilization of end-stage cells.
Terminal differentiation of murine erythroleukemia cells: physical stabilization of end-stage cells.
复制标题
鼠红血症细胞的末端分化：终阶段细胞的物理稳定。
DOI：
10.1083/jcb.93.2.390
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发表时间：
1982-05
影响因子：
7.8
通讯作者：
Housman, D
中科院分区：
文献类型：
--
作者：
Volloch, V;Housman, D
使用鼠红白血病（MEL）细胞系统作为研究终末红细胞分化的体外系统的一个重要限制是不能产生代表体外途径终点的大量细胞。我们在这里表明，未能观察到终末期细胞在体外的一个主要原因是，这些细胞是物理上不稳定的MEL细胞分化的标准培养条件下。通过添加牛血清白蛋白或Ficoll来修改这些培养条件，导致终末期细胞的物理稳定。在这样的培养条件下，观察到形态特征与正常小鼠嗜正染性成红细胞和网织红细胞相似的终末分化MEL细胞的均匀培养物。这些细胞群的物理和生化参数的检查给出的值与小鼠网织红细胞的特征值相似。物理稳定的MEL细胞显示出窄的细胞体积分布，平均值约为100 μ m（3），与小鼠网织红细胞观察到的细胞体积分布相似，而用DMSO但不含稳定剂处理的典型MEL细胞培养物显示出更宽、更异质的细胞体积分布，平均值约为500 μ m（3）。珠蛋白mRNA水平和珠蛋白合成水平达到几乎等于在物理稳定化的MEL细胞的培养物中的小鼠网织红细胞中的那些的值，而未用稳定剂处理的分化培养物达到这些参数的实质上更低的值。我们建议，在体外进行完整的终末红系分化的MEL细胞群体的产生能力将允许控制红系分化过程的终末阶段的分子机制的分析。
An important limitation in the use of the murine erythroleukenia (MEL) cell system as an in vitro system for the study of terminal erythroid differentiation has been the inability to produce significant numbers of cells which represent the end-point of the pathway in vitro. We show here that a major reason for the failure to observe end-stage cells in vitro is that such cells are physically unstable under the standard culture conditions used for MEL cell differentiation. Modification of these culture conditions by the addition of either bovine serum albumin or Ficoll leads to physical stabilization of end-stage cells. Under such culture conditions, uniform cultures of terminally differentiated MEL cells with morphological characteristics similar to those of normal mouse orthochromatophilic erythroblasts and reticulocytes are observed. Examination of physical and biochemical parameters of these cell populations give values which are similar to values characteristic of mouse reticulocytes. A physically stabilized MEL cell shows a narrow cell volume distribution with an average value of approximately 100 mum(3), similar to the cell volume distribution observed for mouse reticulocytes, while a typical MEL cell culture treated with DMSO but without a stabilizing agent exhibits a broader, more heterogeneous cell volume distribution with an average value of approximately 500 mum(3). Globin mRNA levels and levels of globin synthesis reach values almost equal to those in mouse reticulocytes in cultures of physically stabilized MEL cells while differentiating cultures not treated with a stabilizing agent reach substantially lower values for these parameters. We suggest that the ability to produce populations of MEL cells which undergo complete terminal erythroid differentiation in vitro will allow the analysis of the molecular mechanisms which control the terminal stages of the erythroid differentiation process.