| Exit from Pluripotency Assay of Mouse Embryonic Stem Cells
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Author:
Date:
2017-08-20
[Abstract] A novel method to assess the dissolution of the core pluripotency transcription-factor circuit of mouse Embryonic Stem Cells (mESCs) has been developed (Ying et al., 2003; Betschinger et al., 2013). In order to efficiently identify genes essential for the break-down of the pluripotency network in mutant mESCs with proliferation defects, we adapted this ‘exit from pluripotency assay’ (Bodak et al., 2017; Cirera-Salinas et al., 2017). The protocol described here has been successfully applied to several mESC lines and is easily transposable from one laboratory to another.
[摘要] 已经开发了评估小鼠胚胎干细胞（mESCs）的核心多能转录因子电路溶解的新方法（Ying等，2003; Betschinger等，2013）。 为了有效识别具有增殖缺陷的突变体mESCs中多能网络分解所必需的基因，我们调整了这种“多能性测定法”（Bodak等，2017; Cirera-Salinas等，2017）。 这里描述的方案已经成功应用于几个mESC系列，并且可以容易地从一个实验室转座到另一个实验室。
【背景】几十年来，科学家已经尝试确定基因与一般（例如胚胎体）或定向（例如，神经元前体细胞）分化方案的mESCs的分化潜能的机制。最近，发现2i培养基允许在体外俘获天真的干细胞（Ying et al。，2008）。 ...
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| Reprogram Murine Epiblast Stem Cells by Epigenetic Inhibitors
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Author:
Date:
2017-03-05
[Abstract] Pluripotent stem cells in the naïve state are highly useful in regenerative medicine and tissue engineering. A robust reprogramming of the primed murine Epiblast Stem Cells (EpiSCs) to naïve pluripotency is feasible via chemical-only approach. This protocol described a method to reprogram murine EpiSCs by MM-401 treatment, which blocks histone H3K4 methylation by MLL1/KMT2A.
[摘要] 初生状态下的多能干细胞在再生医学和组织工程中非常有用。 通过化学方法，将引发的鼠上皮细胞干细胞（EpiSCs）的鲁棒重新编程成为初始多能性是可行的。 该方案描述了通过MM-401处理重编程小鼠EpiSCs的方法，其通过MLL1 / KMT2A阻断组蛋白H3K4甲基化。
背景 以前关于EpiSC重编程的方案取决于转录因子的遗传操作或信号通路的化学抑制，尽管效率和持续时间不同。 基于最近将MLL1复合物与初始状态联系起来的机制研究，该方案提供了一种直观而有力的方法，通过靶向MLL1介导的H3K4甲基化和随后的转录调节来恢复EpiSC的初始多能性。 重编程效率明显高于以前公布的方法，在两周内达到50％的转化率。
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| Retinal Differentiation of Mouse Embryonic Stem Cells
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Author:
Date:
2016-07-05
[Abstract] Groundbreaking studies from Dr. Yoshiki Sasai’s laboratory have recently introduced novel methods to differentiate mouse and human Embryonic Stem Cells (mESCs and hESCs) into organ-like 3D structures aimed to recapitulate developmental organogenesis programs (Eiraku et al., 2011; Eiraku and Sasai, 2012; Nakano et al., 2012; Kamiya et al., 2011). We took advantage of this method to optimize a 3D protocol to efficiently generate retinal progenitor cells and subsequently retinal neurons in vitro. This culture system provides an invaluable platform both to study early developmental processes and to obtain retinal neurons for transplantation approaches. The protocol described here has been successfully applied to several mouse ESC (including the R1, WD44 and ...
[摘要] 来自Yoshiki Sasai博士的实验室的开创性研究最近已经引入了将小鼠和人胚胎干细胞（mESC和hESC）区分为器官样3D结构的新方法，其旨在重现发育器官发生程序（Eiraku等人， ，2011; Eiraku和Sasai，2012; Nakano等人，2012; Kamiya等人，2011）。 我们利用这种方法优化3D协议以有效地生成视网膜祖细胞和随后视网膜神经元体外。 这种文化系统提供了一个宝贵的平台，既研究早期发展过程，并获得视网膜神经元的移植方法。 这里描述的协议已成功应用于几个鼠标ESC（包括R1，WD44和G4细胞系）和小鼠诱导多能干细胞（iPSCs）线。
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