三种方法对细胞周期诱导效率的比较

Comparison on Three Methods of Cell Cycle Induction

比较两种处理方法对细胞周期调控的影响,本研究用饥饿法,胸腺嘧啶处理法和饥饿法结合胸腺嘧啶法诱导胎儿成纤维细胞进入G1/G0期。诱导两周后,收集3种方法来源的细胞用流式细胞技术、细胞活力检测、real-time PCR和核型分析检查细胞诱导效率、细胞活力、成纤维细胞特异基因的表达水平和核型情况。结果表明,实验组细胞G1/G0期比率显著高于对照组(p<0.05);实验组中,饥饿联合药物诱导法比饥饿法和药物法稍高,但未达显著水准。实验组与对照组细胞核型正常,均为40条染色体,性染色体为XX。细胞活力和显示,对照组比试验组稍高,但差异不显著(p>0.05);FGF4、FGF2及Sox2的基因表达水平在各组间差异不显著(p>0.05)。这些结果表明,实验组3种方法对细胞特性没有显著影响,饥饿结合药物诱导可适当提高细胞诱导效率,为相关研究提供参考。

To investigate the effect of several methods on regulation of cell cycle, the starvation, combination of starvation with TdR and TdR, were used to induce mouse embryonic fibroblasts（MEF） into G1/G0 phase. After 2weeks, the cell was checked by cell viability, cell flow and karyotype analysis. The results showed that rate of G1/G0 phase in treated groups was higher than that in the control group（p〈0.05）; in the treated groups, rate of G1/G0 in combination of starvation with TdR was slightly higher than that in starvation and TdR groups, but no significant difference（p〉0.05）. Karyotypes were normal in treated and control groups, in which, number of chromosomes was40 and gender chromosomes was XX. For cell viability, the viability in control group was slightly higher than treated groups, but which is no significant difference（p〉0.05）. Expression levels of FGF4, FGF2 and Sox2 were not significant difference（p〉0.05）. The results demonstrated that the characteristics of cells was not affected by three methods, but rate of G1/G0 phase of cells could slightly be improved in combination of starvation with TdR, which provided inferences for related researches.