血清饥饿和接触抑制两种G0期同步化方法效果评价

Effects of serum starvation and contact inhibition on the cell cycle G0 synchronization of the human embryonic lung fibroblast

目的 评价血清饥饿和接触抑制两种方法对细胞周期G0期同步化效果，并分析再进入细胞周期各期的时点。方法 利用体外细胞培养技术，采用流式细胞术分析人胚肺成纤维细胞血清饥饿0～48h以及恢复血清培养0～24h内细胞周期分布改变；分析接触抑制3天及传代后24h钿胞周期分布改变。结果 血清饥饿48h后能够达到较好的同步化效果，血清再刺激16h细胞进入细胞周期进程，G1期的细胞在16h后明显减少，24h时达到最低点；S期的细胞在16h后明显增加，于20h左右达到峰值；G2期的细胞在20h时增加明显，24h时达峰值。接触抑制同步化3天后G0达83．36％，传代后，G1期的细胞在13h后逐渐减少，S期的细胞在13h后逐渐增加，于22h时达峰值，G2期的细胞在19h后逐渐增加，25h时达峰值。结论血清饥饿48h或者接触抑制3天均能使体外培养人胚肺成纤维细胞达到G0期同步化状态，恢复血清16h和传代后13h细胞逐渐进入周期。血清饥饿法要优于接触抑制法。

To compare the effects of two different cell cycle GO stage synchronization methods in the human embryonic lung fibroblast and re-entry into the cell cycle. Methods Cell cycle synchronization was achieved by serum starvation and contact inhibition. Which were identified by flow cytometry. Cell morphologic character were also analyzed. Detection cell cycle distribution change after serum starvation（0.5 % serum） for 48h then serum re-stimulation 24h, growth in contact inhibition 72h and subsequent passage 0-24h respectively. Results Serum starvation 48h could well synchronize HELF in GO stage. Re-stimulated by serum 16h cell re-entered cycle and S increased to peak at 20h, G2 increased to peak at 24h. Growth in contact inhibition 72h could synchronize 83.36% cell in GO. Cells re-entered cycle after passage 13h and S increased to peak at 22h, G2 increased to peak at 25h. Conclusion Serum starvation 48h and growth in confluence 72h both can synchronize HELF in GO stage effectively. Cells could re-enter cell cycle 16h after restimulated by serum or 13h after passage respectively. Serum starvation method was better than contact inhibition.