Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
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DOI：
10.1111/j.1476-5381.2010.01197.x
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发表时间：
2011-04-01
影响因子：
7.3
通讯作者：
Sachinidis, A.
中科院分区：
文献类型：
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作者：
Jagtap, S.;Meganathan, K.;Sachinidis, A.
致畸物质在胚胎发育过程中引起不良反应。人胚胎干细胞（human embryonic stem cells，hESC）的多向分化模拟了体外胚胎发育过程。在这里，我们提出了一种在hESC中的转录组学方法，用于监测化合物的特定毒性作用，作为需要大量动物的传统耗时且成本密集的体内试验的替代方案。本研究旨在探讨阿糖胞苷（acytosine arabinoside，Ara-C）对随机分化的人胚胎干细胞（hESCs）的不良影响。亚致死浓度的Ara-C在分化期间的第7天和第14天两个时间点给予。用微阵列评估基因表达，以确定Ara-C存在下的失调转录本。关键词：随机分化的hESC能够产生多谱系标记。低浓度的Ara-C（1 nM）诱导外胚层，并在第14天抑制中胚层。观察到外胚层标志物如MAP 2、TUBB III、PAX 6、TH和NESTIN的诱导，以及中胚层标志物如HAND 2、PITX 2、GATA 5、MYL 4、TNT 2、COL 1A 1和COL 1A 2的抑制。此外，没有观察到细胞凋亡的诱导。基因本体论揭示了与神经元分化和中胚层发育相关的独特的失调生物学过程。通路分析表明，轴突的指导途径失调。结论和implicationsOur结果表明，hESCs与toxicogenomics相结合，提供了一个敏感的体外发育毒性模型作为替代传统的动物实验。
BACKGROUND AND PURPOSETeratogenic substances induce adverse effects during the development of the embryo. Multilineage differentiation of human embryonic stem cells (hESCs) mimics the development of the embryo in vitro. Here, we propose a transcriptomic approach in hESCs for monitoring specific toxic effects of compounds as an alternative to traditional time-consuming and cost-intensive in vivo tests requiring large numbers of animals. This study was undertaken to explore the adverse effects of cytosine arabinoside (Ara-C) on randomly differentiated hESCs.EXPERIMENTAL APPROACHHuman embryonic stem cells were used to investigate the effects of a developmental toxicant Ara-C. Sublethal concentrations of Ara-C were given for two time points, day 7 and day 14 during the differentiation. Gene expression was assessed with microarrays to determine the dysregulated transcripts in presence of Ara-C.KEY RESULTSRandomly differentiated hESCs were able to generate the multilineage markers. The low concentration of Ara-C (1 nM) induced the ectoderm and inhibited the mesoderm at day 14. The induction of ectodermal markers such as MAP2, TUBB III, PAX6, TH and NESTIN was observed with an inhibition of mesodermal markers such as HAND2, PITX2, GATA5, MYL4, TNNT2, COL1A1 and COL1A2. In addition, no induction of apoptosis was observed. Gene ontology revealed unique dysregulated biological process related to neuronal differentiation and mesoderm development. Pathway analysis showed the axon guidance pathway to be dysregulated.CONCLUSIONS AND IMPLICATIONSOur results suggest that hESCs in combination with toxicogenomics offer a sensitive in vitro developmental toxicity model as an alternative to traditional animal experiments.