流式细胞仪分析法检测黑熊成纤维细胞周期同步化处理效果

Effect of synchronization-treated bear fibroblast cells cycle analysed by flow cytometer

分离培养黑熊皮肤成纤维细胞 ,传 3代 ,分别经血清饥饿 2～ 6 d和 10 g/ L Nocodazole处理 2 4 h后 ,用流式细胞仪分析 2个处理组和对照组细胞的周期相 ,以研究黑熊成纤维细胞同步化处理后的细胞周期时相的变化 ,探讨核移植供体细胞的细胞周期对核移植胚胎发育的影响。结果显示 ,血清饥饿组、Nocodazole处理组及对照组的 G0 +G1 期成纤维细胞的百分率分别为 (85 .0 5± 7.0 0 ) % ,(4 5 .9± 3.7) %和 (5 9.3± 6 .7) % ;G2 / M期细胞的百分率分别为 (10 .4± 6 .8) % ,(4 6 .3± 4 .2 ) %及 (2 3.0 5± 1.0 5 ) %。表明血清饥饿法能较好地使黑熊成纤维细胞同步于 G0 和 G1 期 ;Nocodazole处理可以使细胞较大程度同步于 G2 和 M期。

After 3 passages of the primary black bear fibroblast cells,the cells were serum starvated for 2-6 days or treated with 10 g/L Nocodazole for 24 h,then calculation of percentages of the treated cells and the control cells in G 0,G 1 and G 2+M phases of the cell cycle by using flow cytometer was conducted respectively,to investigate the cell cycle characteristics of different treatment cultured fibroblast cells.The results showed that:G 0+G 1 cell percentage of the serum starvation,Nocodazole treatment,and control are (85.05±7.00)%,(45.9±3.7)% and (59.3±6.7)%,respectively.G 2/M are (10.4±6.8)%,(46.3± 4.2)% and (23.05±1.05)%,respectively.Black bear fibroblast cell treated with serum starvation for 5 days contained higher ( P <0.05) percentages of G 0+G 1,whereas cells treated with nocodazole contained higher percentage of G 2+M.