Enrichment of Pluripotent Stem Cell-Derived Hepatocyte-Like Cells by Ammonia Treatment.
Enrichment of Pluripotent Stem Cell-Derived Hepatocyte-Like Cells by Ammonia Treatment.
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DOI：
10.1371/journal.pone.0162693
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Sasaki K
中科院分区：
文献类型：
--
作者：
Tomotsune D;Hirashima K;Fujii M;Yue F;Matsumoto K;Takizawa-Shirasawa S;Yokoyama T;Sasaki K
胚胎干细胞（ESC）和诱导多能干细胞（iPSC）是包括肝脏在内的缺陷器官再生的潜在资源。然而，在这成为现实之前，必须克服一些障碍。分化后保留的未分化细胞具有形成畸胎瘤的潜力。此外，实际应用需要大量的分化细胞，因此分化过程必须是经济的。在这里，我们描述了一种基于DNA微阵列的分化人类多能干细胞的基因表达谱的全球分析。我们确定了六种人类多能干细胞系之间的差异和共性：hESC KhES 1，KhES 2，KhES 3和H1，以及iPSC 201 B7和243 G1。胚状体（EBs）的形成不需要补充诱导因子。EB还表达一些肝脏特异性代谢基因，包括氨代谢酶谷氨酰胺合成酶和氨甲酰磷酸合成酶1。实时荧光定量PCR分析显示，在Lanford培养基中氨处理的EB的肝细胞样分化。DNA微阵列数据的分析表明，肝细胞样细胞是氨处理的细胞中最丰富的群体。此外，未分化的多能干细胞标志物的表达水平急剧降低，表明畸胎瘤形成能力降低。这些结果表明，在缺乏昂贵的诱导因子的情况下，在Lanford培养基中用氨处理EB可能是肝分化的有效诱导剂。
Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are potential resources for the regeneration of defective organs, including the liver. However, some obstacles must be overcome before this becomes reality. Undifferentiated cells that remain following differentiation have teratoma-forming potential. Additionally, practical applications require a large quantity of differentiated cells, so the differentiation process must be economical. Here we describe a DNA microarray-based global analysis of the gene expression profiles of differentiating human pluripotent stem cells. We identified differences and commonalities among six human pluripotent stem cell lines: the hESCs KhES1, KhES2, KhES3, and H1, and the iPSCs 201B7 and 243G1. Embryoid bodies (EBs) formed without requiring supplementation with inducing factors. EBs also expressed some liver-specific metabolic genes including the ammonia-metabolizing enzymes glutamine synthetase and carbamoyl-phosphate synthase 1. Real-time PCR analysis revealed hepatocyte-like differentiation of EBs treated with ammonia in Lanford medium. Analysis of DNA microarray data suggested that hepatocyte-like cells were the most abundant population in ammonia-treated cells. Furthermore, expression levels of undifferentiated pluripotent stem cell markers were drastically reduced, suggesting a reduced teratoma-forming capacity. These results indicate that treatment of EBs with ammonia in Lanford medium may be an effective inducer of hepatic differentiation in absence of expensive inducing factors.
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影响因子：
64.5
作者：
Hanna JH;Saha K;Jaenisch R
通讯作者：
Jaenisch R
影响因子：
3.7
作者：
Tomizawa M;Shinozaki F;Sugiyama T;Yamamoto S;Sueishi M;Yoshida T
通讯作者：
Yoshida T
影响因子：
2.5
作者：
He, Youji;Hakvoort, Theodorus B. M.;Van Roon, Maria A.
通讯作者：
Van Roon, Maria A.
影响因子：
2.3
作者：
Lee SY;Kim HJ;Choi D
通讯作者：
Choi D
影响因子：
4.8
作者：
Behbahan, Iman Saramipoor;Duan, Yuyou;Lam, Alexander;Khoobyari, Shiva;Ma, Xiaocui;Ahuja, Tijess P.;Zern, Mark A.
通讯作者：
Zern, Mark A.