Wittayarat M, Thongphakdee A, Saikhun K, Chatdarong K, Otoi T, Techakumphu M
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Sciences, Chulalongkorn University, Bangkok, Thailand.
Reprod Domest Anim. 2013 Apr;48(2):305-10. doi: 10.1111/j.1439-0531.2012.02149.x. Epub 2012 Jul 27.
This study was examined whether the species of felid affects synchronization accuracy at the G0/G1 stage of the cell cycle and the occurrence of apoptosis by different protocols, such as serum starvation, confluent and roscovitine treatment. Skin fibroblast cells were obtained from the Asian golden cat, marbled cat, leopard and Siamese cat. The cells from each animal were treated with either serum starvation for 1-5 days, cell confluency-contact inhibition for 5 days or roscovitine at various concentrations (7.5-30 μm). Flow cytometric analysis revealed that serum starvation for 3 days provided the highest cell population arrested at the G0/G1 stage, irrespective of the felid species. In all species, 100% confluency gave a significantly higher percentage of cells arrested at the G0/G1 stage compared with the non-treated control cells. The effects of roscovitine treatment and the appropriate concentration on the rates of G0/G1 cells differed among the felid species. Serum starvation for more than 4 days in the marbled cat and Siamese cat and roscovitine treatment with 30 μm in the Asian golden cat and leopard increased the rates of apoptosis. In conclusion, different felid species responded to different methods of cell cycle synchronization. Asian golden cat and Siamese cat fibroblast cells were successfully synchronized to G0/G1 stage using the serum starvation and roscovitine treatment, whereas only confluency-contact inhibition treatment induced cell synchronization in the leopard. Moreover, these three methods did not successfully induce cell synchronization of the marbled cat. These findings may be valuable for preparing their donor cells for somatic cell nuclear transfer in the future.
本研究探讨了猫科动物的种类是否会影响细胞周期G0/G1期的同步化准确性以及不同处理方案（如血清饥饿、汇合培养和罗哌卡因处理）诱导的细胞凋亡情况。从亚洲金猫、云猫、豹和暹罗猫获取皮肤成纤维细胞。将每只动物的细胞分别用血清饥饿处理1 - 5天、汇合培养接触抑制处理5天或用不同浓度（7.5 - 30μm）的罗哌卡因处理。流式细胞术分析显示，无论猫科动物种类如何，血清饥饿3天可使处于G0/G1期停滞的细胞群体比例最高。在所有物种中，100%汇合培养时处于G0/G1期停滞的细胞百分比显著高于未处理的对照细胞。罗哌卡因处理及其合适浓度对G0/G1期细胞比例的影响在不同猫科动物种类中有所不同。云猫和暹罗猫血清饥饿超过4天以及亚洲金猫和豹用30μm罗哌卡因处理会增加细胞凋亡率。总之，不同猫科动物种类对不同的细胞周期同步化方法有不同反应。亚洲金猫和暹罗猫的成纤维细胞通过血清饥饿和罗哌卡因处理成功同步到G0/G1期，而只有汇合培养接触抑制处理能诱导豹的细胞同步化。此外，这三种方法均未成功诱导云猫的细胞同步化。这些发现可能对未来为体细胞核移植制备其供体细胞具有重要价值。
