Abstract：
Fibroblast cycle synchronization in G0/G1 is an essential step for nuclear reprogramming by cloning or induced cells to pluripotency. Considering the diversity among rodents and the ecological and scientific importance of these animals, we compared the contact inhibition, serum starvation, and 10 µM of roscovitine as methods of synchronization of red-rumped agouti fibroblasts. The effects of each protocol were evaluated on the percentage of cycle phase, morphology, viability, and apoptosis levels. The results showed that culturing the cells to serum starvation for 24 h (75.9%), 48 h (81.6%), 72 h (86.2%), 96 h (84.0%), and 120 h (83.7%) yielded a significantly higher percentage of cells arrested in the G0/G1 (P < 0.05) phase than cells not subjected to any cell cycle synchronization method (31.4%). Also, this effect was not different between the times of 48 and 120 h (P > 0.05). A similar response was observed for cells cultured with roscovitine for 12 h (86.9%), 24 h (74.8%), and 48 h (81.7%), with a higher percentage of synchronized cells in G0/G1 compared to cells not submitted to any synchronization treatment (52.2%). Nevertheless, this effect was best evidenced at 12 h (P < 0.05). Also, the contact inhibition for 24-120 h could not synchronize cells in G0/G1, with values ranging from 70.9 to 77.9% (P > 0.05). Moreover, no difference was observed for morphology, viability, and apoptosis levels in any synchronization method (P > 0.05). Therefore, serum starvation is as efficient as roscovitine on cycle synchronization in G0/G1 of red-rumped agouti fibroblasts.
摘要：
G0/G1中的成纤维细胞周期同步是通过克隆或诱导细胞成多能性进行核重编程的必要步骤。考虑到啮齿动物的多样性以及这些动物的生态和科学重要性，我们比较了接触抑制，血清饥饿,和10µM的roscovitine作为红腰刺鼠成纤维细胞的同步方法。每个方案的效果都是根据周期阶段的百分比进行评估的，形态学,生存能力，和凋亡水平。结果表明,培养细胞至血清饥饿24h(75.9%),48小时（81.6%），72小时（86.2%），96小时（84.0%），和120h（83.7％）产生的G0/G1期停滞的细胞百分比（P<0.05）明显高于未进行任何细胞周期同步方法的细胞（31.4％）。此外，48h和120h之间的效果没有差异（P>0.05）。用roscovitine培养12小时的细胞观察到类似的反应（86.9％），24小时（74.8%），和48小时（81.7%），与未接受任何同步治疗的细胞相比，G0/G1中同步细胞的百分比更高（52.2％）。然而，12h时效果最好(P<0.05)。此外，接触抑制24-120h不能使G0/G1细胞同步，值在70.9-77.9％之间（P>0.05）。此外，在形态学上没有观察到差异，生存能力，和凋亡水平在任何同步方法(P>0.05)。因此,血清饥饿与roscovitine一样有效，可以在红Rumpedagouti成纤维细胞的G0/G1中进行周期同步。
