Qiu Xiaoyan, You Haihong, Xiao Xiong, Li Nan, Li Yuemin
Embryo Engineering Laboratory, School of Animal Science and Technology, Southwest University , Chong Qing, P.R. China .
Cell Reprogram. 2017 Feb;19(1):1-9. doi: 10.1089/cell.2016.0030.
The low success rate of animal cloning by somatic cell nuclear transfer (SCNT) is believed to be associated with aberrant epigenetic nuclear reprogramming. It has been demonstrated that treatment with histone deacetylase inhibitors (HDACis) enhances developmental potential of SCNT embryos. Previous studies in many species revealed that treatment of SCNT embryos with trichostatin A (TSA)-an HDACi-significantly enhances the in vitro development of SCNT embryos. In this study, we compared two different SCNT protocols with TSA and investigate, for the first time, the effect of another new HDACi, PXD101 (belinostat), on in vitro development of mouse SCNT embryo. Rates of blastocyst development in mouse SCNT embryos treated with either 5 nM TSA during (6 hours) and after (4 hours) activation (39.1%) or with 50 nM PXD101 during (6 hours) and after (4 hours) activation (40.2%) were significantly higher than those of nontreated SCNT embryos (11.5%) and both treatments also significantly improved the subsequent establishment of NT-ESCs in comparison with the nontreated group (38.1% and 40.9% vs. 11.8%). In conclusion, we optimized the TSA concentration and treatment timing and, for the first time, investigated the effect of PXD101 on mouse development of SCNT embryos and establishment of NT-ESCs.
体细胞克隆技术（SCNT）的低成功率被认为与异常的表观遗传核重编程有关。已有研究表明，使用组蛋白去乙酰化酶抑制剂（HDACis）处理可提高SCNT胚胎的发育潜力。此前在许多物种中的研究发现，用曲古抑菌素A（TSA）——一种HDACi——处理SCNT胚胎，可显著提高其体外发育能力。在本研究中，我们比较了两种不同的SCNT方案与TSA的效果，并首次研究了另一种新型HDACi PXD101（贝利司他）对小鼠SCNT胚胎体外发育的影响。在激活期间（6小时）和激活后（4小时）用5 nM TSA处理的小鼠SCNT胚胎（囊胚发育率为39.1%），以及在激活期间（6小时）和激活后（4小时）用50 nM PXD101处理的小鼠SCNT胚胎（囊胚发育率为40.2%），其囊胚发育率均显著高于未处理的SCNT胚胎（11.5%），并且与未处理组相比，这两种处理还显著提高了随后NT-ESC的建立效率（分别为38.1%和40.9%，而未处理组为11.8%）。总之，我们优化了TSA的浓度和处理时间，并首次研究了PXD101对小鼠SCNT胚胎发育及NT-ESC建立的影响。
