Extended pluripotent stem cells facilitate mouse model generation.
Extended pluripotent stem cells facilitate mouse model generation.
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扩展的多能干细胞促进小鼠模型的生成
DOI：
10.1007/s13238-018-0573-0
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发表时间：
2019-01
期刊：
影响因子：
21.1
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Xiang G;Wang H
1981年建立的小鼠胚胎干（mES）细胞（Evans和考夫曼，1981; Martin，1981）来源于囊胚的内细胞团（ICM），可在体外扩增多次传代，保持正常核型和分化潜能。在引入囊胚后，mES细胞可以分化成所有三个胚层，有助于所有体细胞谱系和种系。1998年，James Thompson从人胚泡的ICM中获得了人胚胎干（hES）细胞（Thomson等人，1998年）。有趣的是，hES细胞具有许多不同于mES细胞的特征，包括形态和维持多能性的信号传导途径（Burdon et al.，2002;福赛斯等人，2002; James等人，2005年）。在2007年，小鼠外胚层干细胞（EpiSC）来源于植入后小鼠胚胎的外胚层（Brons等人，2007; Tesar等人，2007年）。这些小鼠EpiSC具有与小鼠ES细胞不同的分子和功能特性，同时在许多方面类似于人类ES细胞。小鼠ES细胞和EpiSC代表植入前和植入后上胚层的体外对应物，这两个阶段分别定义为幼稚和启动多能性（Nichols和Smith，2009）。传统的人ES细胞类似于小鼠EpiSC作为致敏多能状态。最近，几个小组描述了诱导和维持人ES细胞处于幼稚样状态的培养条件（Chan et al.，2013; Duggal等人，2015; Gafni等人，2013; Takashima等人，2014;泰尼森等人，2014; Valamehr等人，2014; Ware等人，2014），这表明人类多能干细胞也有这两个阶段。虽然多能干细胞可以分化成成体生物体中的所有细胞类型，但幼稚ES细胞和EpiSC都不能促进胚胎外（ExEm）组织，其介导子宫着床和随后的生长胚胎和胎儿的母体营养（Beddington和Robertson，1989）。在2017年，两个小组报道了延伸（或扩增）多能干（EPS）细胞的衍生，其可以在体内产生胚胎和胚胎外谱系（Yang et al.，2017 a，B）。EPS细胞可以有效地来源于早期胚胎，并通过重编程，在人类和小鼠。值得注意的是，一个单一的EPS细胞注入八细胞胚胎可以有助于胚胎本身和滋养外胚层谱系。单细胞转录组分析揭示了EPS细胞中卵裂球特异性签名的富集。mES细胞最重要的应用之一是产生基因敲除小鼠。在这一期的《蛋白质与细胞》杂志上，登实验室的两篇论文（杜等人，2018年; Li等人，2018）显示，与mES细胞相比，EPS细胞在产生小鼠模型方面具有上级优势。Li等表明，EPS细胞在长期培养后具有比ES细胞更好的遗传和表观遗传稳定性。将单个EPS或ES细胞注射到8细胞胚胎中，EPS细胞表现出更好的嵌合贡献能力。他们进一步使用CRISPR-Cas9将人IL 3和IL 6基因敲入小鼠内源基因座。将这些工程化的EPS细胞注射到四倍体胚胎中后，他们能够直接获得IL 3和IL 6基因敲入小鼠，效率为10个注射胚胎中有1只小鼠，而注射ES细胞未能获得任何活产。这些结果表明，小鼠EPS细胞中的基因靶向结合四倍体互补（Nagy et al.，1993）可在2-3个月内有效地建立小鼠模型，但由于目前只有少数小鼠品系可用于ES细胞的衍生，Du等尝试了一种新的方法。
Mouse embryonic stem (mES) cells, established in 1981 (Evans and Kaufman, 1981; Martin, 1981), were derived from the inner cell mass (ICM) of blastocysts and can be expanded in vitro for many passages, maintaining normal karyotype and differentiation potential. Upon introduction into blastocysts, mES cells can differentiate into all three germ layers, contributing to all the somatic lineages and germline. In 1998, James Thompson derived human embryonic stem (hES) cells from the ICM of human blastocysts (Thomson et al., 1998). Intriguingly, hES cells have many characteristics different from mES cells, including morphology and signaling pathway maintaining pluripotency (Burdon et al., 2002; Forsyth et al., 2002; James et al., 2005). In 2007, mouse epiblast stem cells (EpiSCs) were derived from the epiblast of post-implantation mouse embryo (Brons et al., 2007; Tesar et al., 2007). These mouse EpiSCs have distinct molecular and functional properties from mouse ES cells, while resemble human ES cells in many ways. Mouse ES cells and EpiSCs represent the in vitro counterpart of preimplantation and postimplantation epiblast, and these two phases were defined as naïve and primed pluripotency respectively (Nichols and Smith, 2009). The traditional human ES cells are similar to mouse EpiSCs as a primed pluripotent state. Recently, several groups described culture conditions to induce and maintain human ES cells at a naïve-like state (Chan et al., 2013; Duggal et al., 2015; Gafni et al., 2013; Takashima et al., 2014; Theunissen et al., 2014; Valamehr et al., 2014; Ware et al., 2014), suggesting that human pluripotent stem cells also have these two phases. Although pluripotent stem cells can differentiate into all the cell types in an adult organism, neither naïve ES cells nor EpiSCs could contribute to extra-embryonic (ExEm) tissues, which mediate uterine implantation and subsequent maternal nutrition of the growing embryo and fetus (Beddington and Robertson, 1989). In 2017, two groups reported the derivation of extended (or expanded) pluripotent stem (EPS) cells, which could generate both embryonic and extra-embryonic lineages in vivo (Yang et al., 2017a, b). EPS cells could be efficiently derived from early embryos and through reprogramming, both in human and mouse. Remarkably, one single EPS cell injected into eight-cell embryo could contribute to both the embryo proper and the trophectoderm lineages. Single-cell transcriptome analysis revealed enrichment for blastomere-specific signature in EPS cells. One of the most important applications of mES cells is to generate knockout mice. In this issue of Protein & Cell, two papers from Deng lab (Du et al., 2018; Li et al., 2018) showed that, compared to mES cells, EPS cells have superior advantages in generating mouse models. Li et al. showed that EPS cells had genetic and epigenetic stability better than ES cells after long-term culturing. When single EPS or ES cell was injected into eight cell embryos, EPS cells showed much better chimeric contribution capability. They further knocked human IL3 and IL6 genes into mouse endogenous loci using CRISPR-Cas9. After injecting these engineered EPS cells into tetraploid embryos, they were able to derive IL3 and IL6 knock-in mice directly with an efficiency of one mouse out of ten injected embryos, while injecting ES cells failed to obtain any live born. These results showed that gene targeting in mouse EPS cells combined with tetraploid complementation (Nagy et al., 1993) can efficiently produce mouse models in approximately 2–3 months.Since only a few mouse strains are permissive for ES cells derivation, Du and colleagues attempted …
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影响因子：
64.5
作者：
Yang H;Wang H;Shivalila CS;Cheng AW;Shi L;Jaenisch R
通讯作者：
Jaenisch R
影响因子：
5.9
作者：
Valamehr, Bahram;Robinson, Megan;Abujarour, Ramzey;Rezner, Betsy;Vranceanu, Florin;Le, Thuy;Medcalf, Amanda;Lee, Tom Tong;Fitch, Michael;Robbins, David;Flynn, Peter
通讯作者：
Flynn, Peter
影响因子：
64.5
作者：
Takashima Y;Guo G;Loos R;Nichols J;Ficz G;Krueger F;Oxley D;Santos F;Clarke J;Mansfield W;Reik W;Bertone P;Smith A
通讯作者：
Smith A
影响因子：
5.2
作者：
Duggal, Galbha;Warrier, Sharat;Heindryckx, Bjorn
通讯作者：
Heindryckx, Bjorn
影响因子：
64.8
作者：
Brons, I. Gabrielle M.;Smithers, Lucy E.;Vallier, Ludovic
通讯作者：
Vallier, Ludovic