Nishino Koichiro, Arai Yoshikazu, Takasawa Ken, Toyoda Masashi, Yamazaki-Inoue Mayu, Sugawara Tohru, Akutsu Hidenori, Nishimura Ken, Ohtaka Manami, Nakanishi Mahito, Umezawa Akihiro
Laboratory of Veterinary Biochemistry and Molecular Biology, Graduate School of Medicine and Veterinary Medicine/Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Center for Animal Disease Control, University of Miyazaki, Miyazaki, Japan.
Regen Ther. 2018 Sep 1;9:71-78. doi: 10.1016/j.reth.2018.08.002. eCollection 2018 Dec.
Human induced pluripotent stem cells (iPSCs) are established by introducing several reprogramming factors, such as Because of their pluripotency and immortality, iPSCs are considered to be a powerful tool for regenerative medicine. To date, iPSCs have been established all over the world by various gene delivery methods. All methods induced high-quality iPSCs, but epigenetic analysis of abnormalities derived from differences in the gene delivery methods has not yet been performed. Here, we generated genetically matched human iPSCs from menstrual blood cells by using three kinds of vectors, i.e., retrovirus, Sendai virus, and episomal vectors, and compared genome-wide DNA methylation profiles among them. Although comparison of aberrant methylation revealed that iPSCs generated by Sendai virus vector have lowest number of aberrant methylation sites among the three vectors, the iPSCs generated by non-integrating methods did not show vector-specific aberrant methylation. However, the differences between the iPSC lines were determined to be the number of random aberrant hypermethylated regions compared with embryonic stem cells. These random aberrant hypermethylations might be a cause of the differences in the properties of each of the iPSC lines.
人诱导多能干细胞(iPSC)是通过导入几种重编程因子而建立的,例如由于其多能性和永生性,iPSC被认为是再生医学的有力工具。迄今为止,世界各地已通过各种基因递送方法建立了iPSC。所有方法都诱导产生了高质量的iPSC,但尚未对因基因递送方法差异而产生的异常进行表观遗传分析。在这里,我们使用三种载体,即逆转录病毒、仙台病毒和游离型载体,从月经血细胞中生成了基因匹配的人iPSC,并比较了它们之间全基因组DNA甲基化谱。虽然异常甲基化的比较显示,仙台病毒载体产生的iPSC在三种载体中异常甲基化位点数量最少,但非整合方法产生的iPSC未显示出载体特异性异常甲基化。然而,与胚胎干细胞相比,iPSC系之间的差异在于随机异常高甲基化区域的数量。这些随机异常高甲基化可能是每个iPSC系特性差异的原因。