Colomb E, Kopp F, Spyratos F, Martin P M
Laboratoire de Cancérologie Expérimentale, CNRS URA 1175, Faculté de Médecine, Marseille, France.
Cytometry. 1989 May;10(3):263-72. doi: 10.1002/cyto.990100305.
The authors have developed a new methodology for characterizing in situ the cell cycle of human mammary epithelial cell lines. Using a SAMBA 200 cell image processor (scanning cytometry), 15 densitometric and textural parameters were computed on each Feulgen-stained nucleus. Parameters computed from the grey level cooccurrence and run-length section matrices allowed assessment of the chromatin pattern. Multiparametric analysis of data defined: 1) the relative position of each cell; 2) the relative positions of groups of cells, each group corresponding to a definite phase of the cell cycle; and 3) the function of these parameters best separating these phases. Files then were constructed for each phase: G0/G1, S, G2/ and M. Using these three files as a reference to classify cells, it was possible to ascertain the phase of the cell cycle for each cell of a population. The MDA AG human cell line synchronized by mitotic selection was used as a model to develop this method. The criteria used to assign cells to G0/G1, S, or G2 was DNA content. Classification in M phase was achieved by visual identification of mitotic cells. This method was checked on unsynchronized MDA AG and then applied to other human cell lines (MDA MB231, MCF-7, T47D C111). Comparison of results obtained by scanning cytometry and flow cytometry showed the proportion of cells assigned to G0/G1, S, and G2/M by the two methods to be similar. This new method removes some of the limitations of flow cytometry by 1) allowing visual verification of each cell analyzed; 2) lowering the number of cells required for study; 3) discriminating between G2 and M; and 4) preserving cell topography.
作者们开发了一种新方法，用于原位表征人乳腺上皮细胞系的细胞周期。使用SAMBA 200细胞图像处理器（扫描细胞术），对每个福尔根染色的细胞核计算15个光密度和纹理参数。从灰度共生矩阵和游程长度剖面矩阵计算得到的参数可用于评估染色质模式。对数据进行多参数分析可确定：1）每个细胞的相对位置；2）细胞组的相对位置，每组对应细胞周期的一个特定阶段；3）最能区分这些阶段的这些参数的函数。然后为每个阶段构建文件：G0/G1、S、G2和M。以这三个文件为参考对细胞进行分类，就可以确定群体中每个细胞的细胞周期阶段。通过有丝分裂选择同步化的MDA AG人细胞系被用作开发此方法的模型。用于将细胞分配到G0/G1、S或G2期的标准是DNA含量。通过目视识别有丝分裂细胞来实现M期的分类。该方法在未同步化的MDA AG细胞上进行了检验，然后应用于其他人类细胞系（MDA MB231、MCF-7、T47D C111）。扫描细胞术和流式细胞术所得结果的比较表明，两种方法分配到G0/G1、S和G2/M期的细胞比例相似。这种新方法消除了流式细胞术的一些局限性，具体如下：1）允许对每个分析的细胞进行目视验证；2）减少研究所需的细胞数量；3）区分G2期和M期；4）保留细胞拓扑结构。
