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MPSS profiling of human embryonic stem cells

机译:人类胚胎干细胞的MPSS分析

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Background Pooled human embryonic stem cells (hESC) cell lines were profiled to obtain a comprehensive list of genes common to undifferentiated human embryonic stem cells. Results Pooled hESC lines were profiled to obtain a comprehensive list of genes common to human ES cells. Massively parallel signature sequencing (MPSS) of approximately three million signature tags (signatures) identified close to eleven thousand unique transcripts, of which approximately 25% were uncharacterised or novel genes. Expression of previously identified ES cell markers was confirmed and multiple genes not known to be expressed by ES cells were identified by comparing with public SAGE databases, EST libraries and parallel analysis by microarray and RT-PCR. Chromosomal mapping of expressed genes failed to identify major hotspots and confirmed expression of genes that map to the X and Y chromosome. Comparison with published data sets confirmed the validity of the analysis and the depth and power of MPSS. Conclusions Overall, our analysis provides a molecular signature of genes expressed by undifferentiated ES cells that can be used to monitor the state of ES cells isolated by different laboratories using independent methods and maintained under differing culture conditions
机译:背景技术对合并的人类胚胎干细胞(hESC)细胞系进行分析,以获得未分化的人类胚胎干细胞共有的基因的完整列表。结果对汇集的hESC细胞系进行分析以获得人ES细胞共有的基因的全面列表。大约300万个签名标签(签名)的大规模并行签名测序(MPSS)识别出近一万一千个独特的转录本,其中约25%是未表征的或新的基因。通过与公共SAGE数据库,EST文库进行比较,并通过微阵列和RT-PCR进行平行分析,确认了先前鉴定的ES细胞标志物的表达,并鉴定了多个ES细胞未知表达的基因。表达基因的染色体作图无法鉴定主要热点,也无法证实映射到X和Y染色体的基因的表达。与已发布数据集的比较证实了分析的有效性以及MPSS的深度和功效。结论总体而言,我们的分析提供了未分化ES细胞表达的基因的分子标记,可用于监测由不同实验室使用独立方法分离并维持在不同培养条件下的ES细胞的状态

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