Qiu Xiaoyan, Li Nan, Xiao Xiong, Zhang Liang, You Haihong, Li Yuemin
Embryo Engineering Laboratory, College of Animal Science and Technology, Southwest University , Chong Qing, P.R. China .
Cell Reprogram. 2017 Dec;19(6):337-343. doi: 10.1089/cell.2017.0027. Epub 2017 Nov 1.
To improve the cloning efficiency of somatic cell nuclear transfer (SCNT) and to establish nuclear transfer embryonic stem cells (NT-ESCs) reliably, it is necessary to produce high-quality blastocysts derived from mice SCNT embryos. Therefore, the present study aims to investigate an optimal method for mouse SCNT embryo production and NT-ESCs derivation by comparing the effects of two methods: the treatment of histone deacetylase inhibitor PXD101 after SCNT, embryo aggregation and their combination treatment. The results suggest that embryo aggregation at four-cell stage and 50 nM PXD101 treated for 10 hours during and after activation could improve both mouse SCNT embryos' development (PXD101: 40.0% vs. 18.5%; p < 0.05; aggregation: 40.2% vs. 18.5%; p < 0.05) and also enhance the isolation rate of NT-ESCs (PXD101: 38.2% vs. 12.5%; p < 0.05; aggregation: 39.0% vs. 12.5%; p < 0.05). The combination of their treatments had a higher development rate (43.6%) and significantly higher NT-ESCs isolation rate (54.7%), therefore, we concluded that the combination of these two methods (50 nM PXD101 treated for 10 hours after SCNT and then aggregated at four-cell stage) is considered as the optimal way for the in vitro development of SCNT embryo and subsequent NT-ESCs isolation in mice, providing a new approach for the practical improvement of mouse cloning techniques and opening new opportunities to improve cloning efficiencies in other species.
为了提高体细胞核移植（SCNT）的克隆效率并可靠地建立核移植胚胎干细胞（NT-ESCs），有必要从小鼠SCNT胚胎中产生高质量的囊胚。因此，本研究旨在通过比较两种方法的效果来研究小鼠SCNT胚胎生产和NT-ESCs衍生的最佳方法：SCNT后组蛋白脱乙酰酶抑制剂PXD101处理、胚胎聚集及其联合处理。结果表明，四细胞期胚胎聚集以及在激活期间和之后用50 nM PXD101处理10小时，既能提高小鼠SCNT胚胎的发育率（PXD101：40.0%对18.5%；p<0.05；聚集：40.2%对18.5%；p<0.05），又能提高NT-ESCs的分离率（PXD101：38.2%对12.5%；p<0.05；聚集：39.0%对12.5%；p<0.05）。它们的联合处理具有更高的发育率（43.6%）和显著更高的NT-ESCs分离率（54.7%），因此，我们得出结论，这两种方法的联合（SCNT后用50 nM PXD101处理10小时，然后在四细胞期聚集）被认为是小鼠SCNT胚胎体外发育和随后NT-ESCs分离的最佳方法，为实际改进小鼠克隆技术提供了新途径，并为提高其他物种的克隆效率带来了新机遇。
