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A Modular Platform for Differentiation of Human PSCs into All Major Ectodermal Lineages

机译:一种模块化平台,用于将人PSC分化为所有主要的外胚层谱系

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摘要

Directing the fate of human pluripotent stem cells (hPSCs) into different lineages requires variable starting conditions and components with undefined activities, introducing inconsistencies that confound reproducibility and assessment of specific perturbations. Here we introduce a simple, modular protocol for deriving the four main ectodermal lineages from hPSCs. By precisely varying FGF, BMP, WNT, and TGF beta pathway activity in a minimal, chemically defined medium, we show parallel, robust, and reproducible derivation of neuroectoderm, neural crest (NC), cranial placode (CP), and non-neural ectoderm in multiple hPSC lines, on different substrates independently of cell density. We highlight the utility of this system by interrogating the role of TFAP2 transcription factors in ectodermal differentiation, revealing the importance of TFAP2A in NC and CP specification, and performing a small-molecule screen that identified compounds that further enhance CP differentiation. This platform provides a simple stage for systematic derivation of the entire range of ectodermal cell types.
机译:将人类多能干细胞(HPSC)的命运引导到不同的谱系中需要可变的起始条件和组分,其中具有未定义的活动,引入了对特定扰动的混淆可重复性和评估的不一致。在这里,我们介绍了一种简单的模块化协议,用于导出来自HPSC的四个主要的异位谱系。通过在最小化学定义的培养基中精确改变FGF,BMP,WNT和TGFβ途径活性,我们显示神经外胚层,神经嵴(NC),颅骨贴片(CP)和非神经网络的平行,鲁棒和可重复的衍生衍生,以及非神经网络在多个HPSC线中,在不同的基材上独立于细胞密度。我们通过询问TFAP2转录因子在异位分化中的作用,揭示了TFAP2a在NC和CP规范中的重要性,并进行了鉴定进一步增强CP分化的化合物的小分子筛网的重要性。该平台提供了一种简单的阶段,用于系统衍生整体凸粒类型。

著录项

  • 来源
    《Cell stem cell》 |2017年第3期|共19页
  • 作者单位

    Sloan Kettering Inst Ctr Stem Cell Biol New York NY 10065 USA;

    Sloan Kettering Inst Ctr Stem Cell Biol New York NY 10065 USA;

    Sloan Kettering Inst Ctr Stem Cell Biol New York NY 10065 USA;

    Cornell Univ Weill Grad Sch Med Sci New York NY 10065 USA;

    Sloan Kettering Inst Ctr Stem Cell Biol New York NY 10065 USA;

    Cornell Univ Dept Surg Weill Med Coll New York NY 10065 USA;

    Sloan Kettering Inst Ctr Stem Cell Biol New York NY 10065 USA;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
  • 关键词

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