Department for Reproductive Medicine, Ghent-Fertility and Stem cell Team (G-FaST), Ghent University Hospital, 9000 Ghent, Belgium.
Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
Nat Commun. 2017 Apr 21;8:15055. doi: 10.1038/ncomms15055.
Until recently, human embryonic stem cells (hESCs) were shown to exist in a state of primed pluripotency, while mouse embryonic stem cells (mESCs) display a naive or primed pluripotent state. Here we show the rapid conversion of in-house-derived primed hESCs on mouse embryonic feeder layer (MEF) to a naive state within 5-6 days in naive conversion media (NCM-MEF), 6-10 days in naive human stem cell media (NHSM-MEF) and 14-20 days using the reverse-toggle protocol (RT-MEF). We further observe enhanced unbiased lineage-specific differentiation potential of naive hESCs converted in NCM-MEF, however, all naive hESCs fail to differentiate towards functional cell types. RNA-seq analysis reveals a divergent role of PI3K/AKT/mTORC signalling, specifically of the mTORC2 subunit, in the different naive hESCs. Overall, we demonstrate a direct evaluation of several naive culture conditions performed in the same laboratory, thereby contributing to an unbiased, more in-depth understanding of different naive hESCs.
直到最近，人们发现人类胚胎干细胞（hESCs）处于初始多能性状态，而小鼠胚胎干细胞（mESCs）则表现出原始或初始多能性状态。在这里，我们展示了在初始转化培养基（NCM-MEF）中，5-6 天内将内部衍生的初始 hESCs 快速转化为初始状态，在初始人干细胞培养基（NHSM-MEF）中 6-10 天，使用反向转换方案（RT-MEF）则需要 14-20 天。我们进一步观察到在 NCM-MEF 中转化的初始 hESCs 的无偏谱系特异性分化潜力增强，然而，所有初始 hESCs 都无法向功能性细胞类型分化。RNA-seq 分析揭示了 PI3K/AKT/mTORC 信号通路的不同作用，特别是 mTORC2 亚基，在不同初始 hESCs 中的不同作用。总的来说，我们在同一个实验室中直接评估了几种初始培养条件，从而为深入了解不同初始 hESCs 提供了更客观、更深入的理解。
