Single-cell qPCR demonstrates that Repsox treatment changes cell fate from endoderm to neuroectoderm and disrupts epithelial-mesenchymal transition
Single-cell qPCR demonstrates that Repsox treatment changes cell fate from endoderm to neuroectoderm and disrupts epithelial-mesenchymal transition
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DOI：
10.1371/journal.pone.0223724
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发表时间：
2019-10
期刊：
影响因子：
3.7
通讯作者：
Qiuhong Li;Qingsong Huang
中科院分区：
文献类型：
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作者：
Qiuhong Li;Qingsong Huang
一个明确的内胚层细胞谱系是高效产生成熟内胚层衍生物的先决条件，这些内胚层衍生物可以产生器官，如胰腺和肝脏。我们以前曾报道，间充质决定性内胚层细胞的诱导依赖于自分泌转化生长因子-β信号，并且Repsox对转化生长因子-β信号通路的药理阻断破坏了内胚层的规范。最后的内胚层起源于原始条纹，这在很大程度上依赖于转化生长因子-β信号。如果转化生长因子-β途径被Repsox阻断，细胞在原始条纹诱导后的命运到目前为止还不清楚。我们在这里报道，诱导的原始条纹细胞群包含许多T/SOX2共表达的细胞，随后Repsox抑制转化生长因子-β信号转导促进了神经外胚层细胞的命运，这是通过单细胞定量聚合酶链式反应和免疫染色来表征的。Repsox治疗也破坏了上皮向间充质转化的过程，这是最终内胚层分化所固有的过程。我们的发现可能为培养神经前体细胞提供一种新的方法。
A definitive endodermal cell lineage is a prerequisite for the efficient generation of mature endoderm derivatives that give rise to organs, such as the pancreas and liver. We previously reported that the induction of mesenchymal definitive endoderm cells depends on autocrine TGF-β signaling and that pharmacological blockage of TGF-β signaling by Repsox disrupts endoderm specification. The definitive endoderm arises from a primitive streak, which depends largely on TGF-β signaling. If the TGF-β pathway is blocked by Repsox, cell fate after the primitive streak induction is so-far unknown. We report here, that an induced primitive streak cell-population contained many T/SOX2 co-expressing cells, and subsequent inhibition of TGF-β signaling by Repsox promoted neuroectodermal cell fate, which was characterized using single-cell qPCR analysis and immunostaining. The process of epithelial-to-mesenchymal transition, which is inherent to the process of definitive endoderm differentiation, was also disrupted upon Repsox treatment. Our findings may provide a new approach to produce neural progenitors.