Kim Ghangyong, Roy Pantu Kumar, Fang Xun, Hassan Bahia Ms, Cho Jongki
College of Veterinary Medicine, Chungnam National University, Daejeon 34134, Korea.
Xenotransplantation Research Center, College of Medicine, Seoul National University, Seoul 03080, Korea.
J Vet Sci. 2019 May;20(3):e31. doi: 10.4142/jvs.2019.20.e31.
This study examined the effects of a caffeine treatment to improve nuclear reprogramming in porcine cloned embryos. Embryonic development and the expression of genes related to pluripotency (, , , and ) were compared after caffeine supplementation during manipulation at different concentrations (0, 1.25, 2.5, and 5.0 mM) and after varying the delayed activation time (control, 1, 2, and 4 h) after fusion. Caffeine added to media during manipulation produced a higher rate of development to blastocysts in the 1.25 mM group than in the other concentration groups (22.8% vs. 16.1%, 16.2%, and 19.2%; < 0.05). When caffeine was added during the 4 h delayed activation, the 1.25 mM caffeine concentration produced a significantly higher rate of development than those in the other 4 h-activation-delayed caffeine concentration groups (22.4% vs. 9.4%, 14.0%, and 11.1%; < 0.05). On the other hand, no significant improvement over that in the control group was observed when caffeine was supplemented during both the manipulation period and delayed activation period (16.0% vs. 15.2%), respectively. The levels of POU5F1, SOX2, and NANOG expression in blastocysts were significantly higher in the delayed activation caffeine group (4 h, 1.25 mM) than in the control group (1 h, 0 mM; < 0.05). In conclusion, a caffeine treatment at 1.25 mM during delayed activation for 4 h can improve the preimplantation development of porcine somatic cell nuclear transfer embryos by activating nuclear reprogramming.
本研究检测了咖啡因处理对改善猪克隆胚胎细胞核重编程的影响。在操作过程中添加不同浓度(0、1.25、2.5和5.0 mM)的咖啡因,并在融合后改变延迟激活时间(对照、1、2和4小时),比较了胚胎发育情况以及多能性相关基因(、、和)的表达。在操作过程中添加到培养基中的咖啡因,1.25 mM组发育至囊胚的比率高于其他浓度组(22.8% 对16.1%、16.2%和19.2%;<0.05)。在4小时延迟激活期间添加咖啡因时,1.25 mM咖啡因浓度组的发育率显著高于其他4小时延迟激活咖啡因浓度组(22.4% 对9.4%、14.0%和11.1%;<0.05)。另一方面,在操作期和延迟激活期均添加咖啡因时,与对照组相比未观察到显著改善(16.0% 对15.2%)。延迟激活咖啡因组(4小时,1.25 mM)囊胚中POU5F1、SOX2和NANOG的表达水平显著高于对照组(1小时,0 mM;<0.05)。总之,在延迟激活4小时期间使用1.25 mM咖啡因处理可通过激活细胞核重编程来改善猪体细胞核移植胚胎的着床前发育。