Comparative proteome and transcriptome analyses of embryonic stem cells during embryoid body-based differentiation
Comparative proteome and transcriptome analyses of embryonic stem cells during embryoid body-based differentiation
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DOI：
10.1002/pmic.200900003
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发表时间：
2009-11-01
期刊：
影响因子：
3.4
通讯作者：
Salekdeh, Ghasem Hosseini
中科院分区：
文献类型：
--
作者：
Fathi, Ali;Pakzad, Mohammad;Salekdeh, Ghasem Hosseini
胚胎干细胞基因表达分析将有助于揭示或进一步确定参与自我更新和多能性维持的信号通路和分子机制。我们采用了一种基于双向电泳的蛋白质组学方法来分析人胚胎干细胞系Royan H5，在未分化的细胞和不同阶段的自发分化（第3，6，12和20天）的胚状体形成。在凝胶上可重复检测的945种蛋白质中，96个点的表达在分化过程中发生了变化。使用MS，鉴定了87种ESC相关蛋白，包括参与细胞增殖、细胞凋亡、转录、翻译、mRNA加工和蛋白质折叠的几种蛋白。还使用微阵列分析了Royan H5分化伴随的转录变化。我们开发了一个全面的数据集，显示使用人ESC系在体外模拟原肠胚形成和器官发生。我们的研究结果表明，蛋白质组学和转录组学数据是互补的，而不是重复的。虽然在分化过程中的许多基因的调控只在转录水平上观察到，几个蛋白质的调制，揭示了只有通过蛋白质组分析。
Gene expression analyses of embryonic stem cells (ESCs) will help to uncover or further define signaling pathways and molecular mechanisms involved in the maintenance of self-renewal and pluripotency. We employed a 2-DE-based proteomics approach to analyze human ESC line, Royan H5, in undifferentiated cells and different stages of spontaneous differentiation (days 3, 6, 12, and 20) by embryoid body formation. Out of 945 proteins reproducibly detected on gels, the expression of 96 spots changed during differentiation. Using MS, 87 ESC-associated proteins were identified including several proteins involved in cell proliferation, cell apoptosis, transcription, translation, mRNA processing, and protein folding. Transcriptional changes accompanying differentiation of Royan H5 were also analyzed using microarrays. We developed a comprehensive data set that shows the use of human ESC lines in vitro to mimic gastrulation and organogenesis. Our results showed that proteomics and transcriptomics data are complementary rather than duplicative. Although regulation of many genes during differentiation were observed only at transcript level, modulation of several proteins was revealed only by proteome analysis.