Gene expression in human embryonic stem cell lines: unique molecular signature
Gene expression in human embryonic stem cell lines: unique molecular signature
复制标题
DOI：
10.1182/blood-2003-09-3314
复制
发表时间：
2004-04-15
期刊：
影响因子：
20.3
通讯作者：
Puri, RK
中科院分区：
文献类型：
--
作者：
Bhattacharya, B;Miura, T;Puri, RK
人胚胎干 (huES) 细胞具有分化成多种细胞谱系的能力，并有可能为许多治疗用途提供分化细胞的来源。然而，人们对huES细胞的分化机制和细胞发育调控因素知之甚少。我们使用含有 16 659 个七十碱基对寡核苷酸的高质量微阵列来检查 11 种可用 huES 细胞系中 6 种的基因表达。将表达与来自多个组织的汇集 RNA（通用 RNA）进行比较，并鉴定了 huES 细胞中富集的基因。所有6个细胞系均表达未分化状态的多个标记，并且在基因表达方面具有显着的同源性（总体相似性系数> 0.85）。确定了 92 个基因的常见子集，包括 Nanog、GTCM-1、连接蛋白 43 (GJA1)、oct-4 和 TDGF1 (cripto)。基因表达通过多种技术得到证实，包括与数据库比较、逆转录酶-聚合酶链式反应、聚焦cDNA微阵列和免疫细胞化学。与已发表的“sternness”基因的比较显示出有限的重叠，表明与其他干细胞群体几乎没有相似之处。几个新的 ES 细胞特异性表达序列标签被鉴定并映射到人类基因组。这些结果代表了未分化 huES 细胞的首次详细表征，并提供了一组独特的标记来分析和更好地了解 huES 细胞的生物学特性。
Human embryonic stem (huES) cells have the ability to differentiate into a variety of cell lineages and potentially provide a source of differentiated cells for many therapeutic uses. However, little is known about the mechanism of differentiation of huES cells and factors regulating cell development. We have used high-quality microarrays containing 16 659 seventy-base pair oligonucleotides to examine gene expression in 6 of the 11 available huES cell lines. Expression was compared against pooled RNA from multiple tissues (universal RNA) and genes enriched in huES cells were identified. All 6 cell lines expressed multiple markers of the undifferentiated state and shared significant homology in gene expression (overall similarity coefficient > 0.85). A common subset of 92 genes was identified that included Nanog, GTCM-1, connexin 43 (GJA1), oct-4, and TDGF1 (cripto). Gene expression was confirmed by a variety of techniques including comparison with databases, reverse transcriptase-polymerase chain reaction, focused cDNA microarrays, and immunocytochemistry. Comparison with published "sternness" genes revealed a limited overlap, suggesting little similarity with other stem cell populations. Several novel ES cell-specific expressed sequence tags were identified and mapped to the human genome. These results represent the first detailed characterization of undifferentiated huES cells and provide a unique set of markers to profile and better understand the biology of huES cells.