Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK; PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan.
Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
Cell. 2014 Sep 11;158(6):1254-1269. doi: 10.1016/j.cell.2014.08.029.
Current human pluripotent stem cells lack the transcription factor circuitry that governs the ground state of mouse embryonic stem cells (ESC). Here, we report that short-term expression of two components, NANOG and KLF2, is sufficient to ignite other elements of the network and reset the human pluripotent state. Inhibition of ERK and protein kinase C sustains a transgene-independent rewired state. Reset cells self-renew continuously without ERK signaling, are phenotypically stable, and are karyotypically intact. They differentiate in vitro and form teratomas in vivo. Metabolism is reprogrammed with activation of mitochondrial respiration as in ESC. DNA methylation is dramatically reduced and transcriptome state is globally realigned across multiple cell lines. Depletion of ground-state transcription factors, TFCP2L1 or KLF4, has marginal impact on conventional human pluripotent stem cells but collapses the reset state. These findings demonstrate feasibility of installing and propagating functional control circuitry for ground-state pluripotency in human cells.
目前的人类多能干细胞缺乏调控小鼠胚胎干细胞（ESC）基础状态的转录因子网络。在这里，我们报告称，短期表达两个组件，NANOG 和 KLF2，足以点燃网络的其他元素并重置人类多能状态。ERK 和蛋白激酶 C 的抑制作用维持着一个独立于转染基因的重布线状态。重置细胞持续自我更新，无需 ERK 信号，表型稳定，染色体完整。它们在体外分化，并在体内形成畸胎瘤。代谢被重新编程，与 ESC 中的线粒体呼吸激活一样。DNA 甲基化显著降低，多个细胞系的转录组状态全面重新调整。基础状态转录因子 TFCP2L1 或 KLF4 的耗竭对传统的人类多能干细胞影响不大，但会破坏重置状态。这些发现证明了在人类细胞中安装和传播基础状态多能性功能控制回路的可行性。
