六种哺乳动物细胞系染色体制片条件优化研究

Optimization of Chromosome Production Conditions for Six Mammalian Cell Lines

关键词:  ［］胞核学检验  染色体制片  条件优化  传代细胞系

Key words:  karyotype analysis, Chromosome preparation  Condition optimization  cell lines

摘要:

［］为了提高胞核学检验的检查效率,采用6种具有种属代表性的常用传代细胞系对影响检验的关键步骤(染色体制片条件)进行优化,并对这六种传代细胞系的染色体的数目进行了统计。本实验以CHO、BHK-21、ST、OA3.Ts、MDBK、RK-13六种疫苗生产中常用的传代细胞系做为材料,对影响染色体制片效果几个主要条件(秋水仙素浓度、KCL浓度及低渗处理时间)进行了优化,并对这六种细胞染色体数目分布进行了检查。结果发现,在细胞用25cm_²细胞培养瓶1:2比例传代24-48 h时,秋水仙素浓度为1 ug/ml,处理60 min后处在中期分裂相的细胞最多,视野中染色体长度适中；用0.4 %KCL溶液低渗处理40 min,细胞中染色体在载玻片上形态最佳,分散度最好。优化后的染色体制片条件,可适用于多种细胞的染色体制片,结果重复性好,染色体形态和分散度均满足胞核学检验的要求,为胞核学检验的流程标准化提供了有益参考。

Abstract:

In order to improve the efficiency of karyotype analysis,six cell lines was used to optimized the main conditions affecting the chromosome production in karyotype analysis, and the number of chromosomes in these six cell lines was counted. Mammalian cell lines CHO, BHK-21, ST, OA3.Ts, MDBK and RK-13 were used as materials to optimize the main conditions (colchicine concentration,KCL solution concentration and low osmotic treatment time) affecting the effect of chromosome slicing in cytological examination. The chromosome number distribution of six cell lines was examined. It was showed that when the cells were subcultured in 25cm_² cell culture flask at a ratio of 1:2 for 24-48 hours, colchicine concentration is 1ug/ml,after 60 minutes of treatment, the cells in the metaphase phase were the most abundant. Chromosome lengths in the field of vision were moderate. Chromosome morphology in the cells was the best when treated with 0.4% KCL solution for 40 minutes. The dispersion is the best. The optimized condition of chromosome production can be applied to many kinds of cells. The results have good repeatability, and the chromosome morphology and dispersion meet the requirements of the karyotype analysis, which provides a useful reference for the standardization of the process of the karyotype analysis.