Stepwise conversion methods between ground states pluripotency from naive to primed.
Stepwise conversion methods between ground states pluripotency from naive to primed.
复制标题
基态多能性从幼稚到引发的逐步转换方法。
DOI：
10.1016/j.bbrc.2021.07.097
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发表时间：
2021
影响因子：
3.1
通讯作者：
Wu J.
中科院分区：
文献类型：
--
作者：
Okamura D;Chikushi M;Chigi Y;Shiogai N;Jafar S;Wu J.
多能干细胞（Pluripotent stem cells，PSC）是体内多能性连续体的体外适应，并且可以广泛地分为具有植入前上胚层特征的幼稚状态和类似于原肠胚形成前上胚层的致敏状态。幼稚PSC和致敏PSC的细胞和分子特征不同，例如，维持未分化状态的分子机制。幼稚至致敏PSC转变提供了一种易于处理的体外模型来研究体内多能性发育。我们之前开发了一种方案，该方案通过在含有FGF 2和Wnt信号传导抑制剂（IWR 1）的培养条件（F/R1条件）下培养分离的植入后小鼠上胚层，使得能够高效（100%）和同质地衍生基态的致敏的上胚层干细胞（rsEpiSC）。基于F/R1条件，本研究开发了三种从小鼠胚胎干细胞（2 i/LIF条件）诱导rsEpiSCs的方法。我们发现，逐步的方法，但不是直接的，是有效的真正的rsEpiSCs从小鼠胚胎干细胞转化。总之，我们建立了一个强大的和有效的基态的幼稚到引发PSC转换策略，这将有助于研究参与体内多能性进展的遗传，表观遗传和代谢过程。
Pluripotent stem cells (PSCs) are in vitro adaptations of in vivo pluripotency continuum and can be broadly classified into naïve state characteristic of pre-implantation epiblast and primed state resembling peri-gastrulation epiblasts. Naïve and primed PSCs differ in their cellular and molecular characteristics, e.g., molecular mechanisms for maintaining undifferentiated state. Naïve-to-primed PSC transition provides a tractable in vitro model to study pluripotency development in vivo. We previously developed a protocol that enabled high-efficient (100%) and homogenous derivation of ground state of primed epiblast stem cells (rsEpiSCs) by culturing the isolated post-implantation mouse epiblast under the culture condition containing FGF2 and a Wnt signaling inhibitor (IWR1) (F/R1 condition). Based on F/R1 condition, in this study, we developed three naïve-to-primed conversion methods for generating rsEpiSCs from naïve ground state of mouse ESCs (2i/LIF condition). We found that stepwise methods, but not directly, were effective for bona fide rsEpiSCs conversion from mouse ESCs. In sum, we established a robust and efficient ground states of naïve-to-primed PSC conversion strategy that will facilitate the study of genetic, epigenetic and metabolic processes involved in pluripotency progression in vivo.