Goissis M D, Caetano H V A, Marques M G, de Barros F R O, Feitosa W B, Milazzotto M P, Binelli M, Assumpção M E O A, Visintin J A
Laboratory of In vitro Fertilization, Cloning and Animal Transgenesis, Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, Brazil.
Reprod Domest Anim. 2007 Dec;42(6):660-3. doi: 10.1111/j.1439-0531.2006.00839.x.
Arrest of cells in G0/G1 cell cycle phase is desired for nuclear transfer procedures. Serum starvation and cell cycle inhibitors are different ways to induce synchronization of the cell cycle. This study investigated the effects of serum starvation and cycloheximide (CHX) on the cell cycle of low (5th) and high (15th) passages fetal porcine fibroblasts. Cell cycle phases were determined using fluorescent activated cell sorting. Fifth passage fibroblast cultures had higher (p < 0.05) proportion of cells in G0/G1 only after 72 h of serum starvation (77.60 +/- 0.65) when compared with non-starved cells (71.44 +/- 1.88). Serum starvation for all periods tested induced an increase (p < 0.05) on proportion of cells in G0/G1 on the 15th passage. No significant differences were observed on the 5th passage cultures exposed to CHX, although, on the 15th passage an increase on proportion of cells was observed after all periods of exposure (p < 0.05). These data indicates that high passage cells in vitro are more susceptible to serum starvation and CHX G0/G1 synchronization.
在核移植过程中，需要使细胞停滞于G0/G1细胞周期阶段。血清饥饿和细胞周期抑制剂是诱导细胞周期同步化的不同方法。本研究调查了血清饥饿和环己酰亚胺（CHX）对低代（第5代）和高代（第15代）胎猪成纤维细胞细胞周期的影响。使用荧光激活细胞分选法确定细胞周期阶段。与未饥饿细胞（71.44±1.88）相比，仅在血清饥饿72小时后，第5代成纤维细胞培养物中处于G0/G1期的细胞比例更高（p<0.05）（77.60±0.65）。在所有测试时间段，血清饥饿均使第15代细胞中处于G0/G1期的细胞比例增加（p<0.05）。在第5代培养物中，暴露于CHX后未观察到显著差异，尽管在第15代培养物中，在所有暴露时间段后均观察到细胞比例增加（p<0.05）。这些数据表明，体外高代细胞对血清饥饿和CHX诱导的G0/G1同步化更敏感。
