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Human Amniotic Epithelial Cells are Reprogrammed More Efficiently by Induced Pluripotency than Adult Fibroblasts

机译:与成人成纤维细胞相比诱导多能性能更有效地重编程人羊膜上皮细胞

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摘要

Cellular reprogramming from adult somatic cells into an embryonic cell–like state, termed induced pluripotency, has been achieved in several cell types. However, the ability to reprogram human amniotic epithelial cells (hAECs), an abundant cell source derived from discarded placental tissue, has only recently been investigated. Here we show that not only are hAECs easily reprogrammed into induced pluripotent stem cells (AE-iPSCs), but hAECs reprogram faster and more efficiently than adult and neonatal somatic dermal fibroblasts. Furthermore, AE-iPSCs express higher levels of NANOG and OCT4 compared to human foreskin fibroblast iPSCs (HFF1-iPSCs) and express decreased levels of genes associated with differentiation, including NEUROD1 and SOX17, markers of neuronal differentiation. To elucidate the mechanism behind the higher reprogramming efficiency of hAECs, we analyzed global DNA methylation, global histone acetylation, and the mitochondrial DNA A3243G point mutation. Whereas hAECs show no differences in global histone acetylation or mitochondrial point mutation accumulation compared to adult and neonatal dermal fibroblasts, hAECs demonstrate a decreased global DNA methylation compared to dermal fibroblasts. Likewise, quantitative gene expression analyses show that hAECs endogenously express OCT4, SOX2, KLF4, and c-MYC, all four factors used in cellular reprogramming. Thus, hAECs represent an ideal cell type for testing novel approaches for generating clinically viable iPSCs and offer significant advantages over postnatal cells that more likely may be contaminated by environmental exposures and infectious agents.
机译:已从几种体细胞中完成了从成年体细胞重编程为胚胎细胞样状态(称为诱导多能性)的过程。然而,重新编程人羊膜上皮细胞(hAECs)的能力,hAECs是从废弃胎盘组织获得的丰富细胞来源,只是在最近才进行了研究。在这里,我们显示不仅hAECs易于重编程为诱导性多能干细胞(AE-iPSCs),而且hAECs的重编程比成人和新生儿的体细胞真皮成纤维细胞更快,更有效。此外,与人包皮成纤维细胞iPSC(HFF1-iPSC)相比,AE-iPSCs表达更高的NANOG和OCT4水平,并表达与分化相关的基因,包括神经元分化的标志物NEUROD1和SOX17,水平降低。为了阐明hAEC更高的重编程效率背后的机制,我们分析了整体DNA甲基化,整体组蛋白乙酰化和线粒体DNA A3243G点突变。与成人和新生儿真皮成纤维细胞相比,hAECs在整体组蛋白乙酰化或线粒体点突变积累方面无差异,而与真皮成纤维细胞相比,hAECs却显示出整体DNA甲基化降低。同样,定量基因表达分析表明,hAEC可内源表达OCT4,SOX2,KLF4和c-MYC,这是细胞重编程中使用的所有四个因素。因此,hAECs代表了一种理想的细胞类型,可用于测试产生临床上可行的iPSC的新方法,并且比出生后的细胞具有明显的优势,而后者更可能受到环境暴露和传染因子的污染。

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