Li Hui, Gao Shuai, Huang Hua, Liu Wenqiang, Huang Huanwei, Liu Xiaoyu, Gao Yawei, Le Rongrong, Kou Xiaochen, Zhao Yanhong, Kou Zhaohui, Li Jia, Wang Hong, Zhang Yu, Wang Hailin, Cai Tao, Sun Qingyuan, Gao Shaorong, Han Zhiming
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.
University of Chinese Academy of Sciences, Chinese Academy of Science, Beijing, People's Republic of China.
Oncotarget. 2017 Jul 18;8(29):47344-47355. doi: 10.18632/oncotarget.17185.
Somatic cell nuclear transfer and transcription factor mediated reprogramming are two widely used techniques for somatic cell reprogramming. Both fully reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells hold potential for regenerative medicine, and evaluation of the stem cell pluripotency state is crucial for these applications. Previous reports have shown that the Dlk1-Dio3 region is associated with pluripotency in induced pluripotent stem cells and the incomplete somatic cell reprogramming causes abnormally elevated levels of genomic 5-methylcytosine in induced pluripotent stem cells compared to nuclear transfer embryonic stem cells and embryonic stem cells. In this study, we compared pluripotency associated genes Rian and Gtl2 in the Dlk1-Dio3 region in exactly syngeneic nuclear transfer embryonic stem cells and induced pluripotent stem cells with same genomic insertion. We also assessed 5-methylcytosine and 5-hydroxymethylcytosine levels and performed high-throughput sequencing in these cells. Our results showed that Rian and Gtl2 in the Dlk1-Dio3 region related to pluripotency in induced pluripotent stem cells did not correlate with the genes in nuclear transfer embryonic stem cells, and no significant difference in 5-methylcytosine and 5-hydroxymethylcytosine levels were observed between fully and partially reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells. Through syngeneic comparison, our study identifies for the first time that Grb10 is associated with the pluripotency state in nuclear transfer embryonic stem cells.
体细胞核移植和转录因子介导的重编程是体细胞重编程中两种广泛使用的技术。完全重编程的核移植胚胎干细胞和诱导多能干细胞在再生医学中都具有潜力，而评估干细胞的多能性状态对于这些应用至关重要。先前的报道表明，Dlk1-Dio3区域与诱导多能干细胞的多能性相关，并且与核移植胚胎干细胞和胚胎干细胞相比，不完全的体细胞重编程会导致诱导多能干细胞中基因组5-甲基胞嘧啶水平异常升高。在本研究中，我们比较了具有相同基因组插入的同基因核移植胚胎干细胞和诱导多能干细胞中Dlk1-Dio3区域中与多能性相关的基因Rian和Gtl2。我们还评估了这些细胞中的5-甲基胞嘧啶和5-羟甲基胞嘧啶水平，并进行了高通量测序。我们的结果表明，诱导多能干细胞中与多能性相关的Dlk1-Dio3区域中的Rian和Gtl2与核移植胚胎干细胞中的基因不相关，并且在完全和部分重编程的核移植胚胎干细胞与诱导多能干细胞之间未观察到5-甲基胞嘧啶和5-羟甲基胞嘧啶水平的显著差异。通过同基因比较，我们的研究首次确定Grb10与核移植胚胎干细胞的多能性状态相关。
