Mamaeva S E
Tsitologiia. 1996;38(8):787-814.
Numerous personal and literary data on the karyotypic variation of cell lines during their establishing and long-term culturing have been reviewed. A new notion about karyotypic evolutionary pathways of cells in culture is presented. A detailed original approach to cytogenetic study of permanent cell lines is given, which allows, via karyotype reconstruction, to obtain finally a new karyotypic characteristics-the generalized reconstructed karyotype (GRK). Application of the cytogenetic approach as a criterion for the control of authenticity, purity and stability of cell lines is discussed. The cell line analysis by means of GRK elicited that the cell line evolution in vitro passes through two stages: a stage of establishment, and a stage of stabilization, both differing in karyotypic variability of cell populations and clonal selection in culture. The data indicate that it is important in experiments to utilize cell lines being in the stage of stabilization and to characterize chromosomally the cell line at the same passage when it is used. Above all, a comparison of karyotypic variations of tumor and leukemic cells in vitro and in vivo has revealed their common karyotypic evolution regularities (a nonrandom character of numerical and structural chromosome changes and the loss of one of the sex chromosomes) and the karyotypic evolutionary regularities characteristic solely of cells in culture. The main of these being a balanced chromosome set in the cell population as a whole and obligatory retention of diploidy in all chromosomes of the normal set by the majority of human and animal cell lines. It has been revealed that no less than two homologs of each autosome are present in cells of at least 85% of examined lines. Other cell lines (at least 15%) of a generally neurogenic origin are shown to be notable by keeping partial or complete monosomies on autosomes throughout the long-term culturing. Peculiarities of the karyotypic evolution of the latter are regarded in detail in addition to the data on the expression of oncogenes and other growth-associated genes in their cells. It is suggested that there are three main compensatory mechanisms through which cell lines with autosomal monosomies may maintain the vitality in culture: polyploidization of initial cell clones, oncogene amplification, mainly of the myc-family oncogenes, and fragmentary or complete extracopying of several autosomes. In summary, perspective of cell line cytogenetics as a field of biology of the cell in culture is discussed.
本文综述了众多关于细胞系在建立和长期培养过程中核型变异的个人及文献资料。提出了一种关于培养细胞核型进化途径的新观点。给出了一种针对永久细胞系细胞遗传学研究的详细原始方法,该方法通过核型重建,最终能够获得一种新的核型特征——广义重建核型(GRK)。讨论了将细胞遗传学方法作为控制细胞系真实性、纯度和稳定性标准的应用。通过GRK对细胞系进行分析表明,体外细胞系的进化经历两个阶段:建立阶段和稳定阶段,这两个阶段在细胞群体的核型变异性以及培养中的克隆选择方面存在差异。数据表明,在实验中使用处于稳定阶段的细胞系并在使用时对同一传代的细胞系进行染色体特征描述非常重要。最重要的是,对肿瘤细胞和白血病细胞在体外和体内的核型变异进行比较,揭示了它们共同的核型进化规律(染色体数目和结构变化的非随机特征以及一条性染色体的丢失)以及仅在培养细胞中特有的核型进化规律。其中主要规律是整个细胞群体中染色体组的平衡以及大多数人和动物细胞系在正常染色体组的所有染色体中强制保留二倍体。研究发现,至少85%的检测细胞系中每个常染色体至少有两个同源染色体。其他通常起源于神经源性的细胞系(至少15%)在长期培养过程中,常染色体上保持部分或完全单体状态,这一点很显著。除了这些细胞中癌基因和其他生长相关基因表达的数据外,还详细探讨了后者核型进化的特点。研究表明,具有常染色体单体的细胞系可能通过三种主要的补偿机制在培养中维持活力:初始细胞克隆的多倍体化、主要是myc家族癌基因的癌基因扩增以及几条常染色体的片段性或完全额外复制。总之,讨论了细胞系细胞遗传学作为培养细胞生物学领域的前景。