胸腺嘧啶核苷诱导肝癌HepG2细胞同步化

Synchronization of HepG2 cells induced by TdR

目的探讨胸腺嘧啶核苷(TdR)诱导肝癌HepG2细胞同步化的方法。方法于对数生长期的HepG2细胞中加入含终浓度为2.5mmoL/L的TdR培养28h后,PBS洗除TdR,加入新鲜血清培养基,此时记为0时刻,分别继续培养0、2、3、4、5、6、7、8、9、12、18、24、28h,收集细胞,同时实验设立对照组,采用流式细胞术检测细胞周期。结果分别于去除TdR后培养4、8、24h获得78.1%的S期细胞、67.2%的G2/M期细胞、86.3%的G1期细胞。结论终浓度为2.5mmoL/L的TdR处理肝癌HepG2细胞28h,再以新鲜培养基培养不同时间,可以获得同步化效果较好的S、G2/M和G1期细胞。

Objective To study method of synchronization in HepG2 cells induced by thymidine （TdR）. Methods After HepG2 cells in the logarithm period were treated by 2.5 mmol/L TdR for 28 hours, the cells were washed twice using PBS to remove TdR and cultured for 0, 2, 3, 4, 5, 6, 7, 8, 9, 12, 18, 24 h continually. Cells were collected and cell cycle was detected by flow cytometry. Control cells were compared in experiment. Results 78. 1% S-phase cells, 67.2% G2/M-phase cells and 86.3 % G1-phase cells were obtained respectively after cells were removed from TdR and cultured for 4, 8, 24 h continually. Conclusions Cells treated by 2.5 mmol/L TdR for 28 hours and then cultured for different time in fresh culture medium shows a good synchronization at G1-phase, S-phase, G2/M- phase.