Yu Yang, Chang Liang, Zhao Hongcui, Li Rong, Fan Yong, Qiao Jie
1] Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China [2] Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, 100191, China [3] Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing, 100191, China.
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China.
Sci Rep. 2015 May 12;5:10114. doi: 10.1038/srep10114.
Human pluripotent stem cells, including cloned embryonic and induced pluripotent stem cells, offer a limitless cellular source for regenerative medicine. However, their derivation efficiency is limited, and a large proportion of cells are arrested during reprogramming. In the current study, we explored chromosome microdeletion/duplication in arrested and established reprogrammed cells. Our results show that aneuploidy induced by somatic cell nuclear transfer technology is a key factor in the developmental failure of cloned human embryos and primary colonies from implanted cloned blastocysts and that expression patterns of apoptosis-related genes are dynamically altered. Overall, ~20%-53% of arrested primary colonies in induced plurpotent stem cells displayed aneuploidy, and upregulation of P53 and Bax occurred in all arrested primary colonies. Interestingly, when somatic cells with pre-existing chromosomal mutations were used as donor cells, no cloned blastocysts were obtained, and additional chromosomal mutations were detected in the resulting iPS cells following long-term culture, which was not observed in the two iPS cell lines with normal karyotypes. In conclusion, aneuploidy induced by the reprogramming process restricts the derivation of pluripotent stem cells, and, more importantly, pre-existing chromosomal mutations enhance the risk of genome instability, which limits the clinical utility of these cells.
人类多能干细胞,包括克隆胚胎干细胞和诱导多能干细胞,为再生医学提供了无限的细胞来源。然而,它们的诱导效率有限,并且很大一部分细胞在重编程过程中停滞。在本研究中,我们探究了停滞的和已建立的重编程细胞中的染色体微缺失/重复情况。我们的结果表明,体细胞核移植技术诱导的非整倍体是克隆人类胚胎以及植入的克隆囊胚来源的原代集落发育失败的关键因素,并且凋亡相关基因的表达模式发生了动态变化。总体而言,诱导多能干细胞中约20%-53%停滞的原代集落显示非整倍体,并且所有停滞的原代集落中P53和Bax均上调。有趣的是,当使用具有预先存在的染色体突变的体细胞作为供体细胞时,未获得克隆囊胚,并且在长期培养后的所得诱导多能干细胞中检测到额外的染色体突变,而在两个核型正常的诱导多能干细胞系中未观察到这种情况。总之,重编程过程诱导的非整倍体限制了多能干细胞的诱导,更重要的是,预先存在的染色体突变增加了基因组不稳定的风险,这限制了这些细胞的临床应用。