Comparison of the gene expression profile of undifferentiated human embryonic stem cell lines and differentiating embryoid bodies.
Comparison of the gene expression profile of undifferentiated human embryonic stem cell lines and differentiating embryoid bodies.
复制标题
DOI：
10.1186/1471-213x-5-22
复制
发表时间：
2005-10-05
影响因子：
--
通讯作者：
Puri RK
中科院分区：
文献类型：
--
作者：
Bhattacharya B;Cai J;Luo Y;Miura T;Mejido J;Brimble SN;Zeng X;Schulz TC;Rao MS;Puri RK
胚胎干细胞（hESC）分化的分子途径的鉴定对于基于干细胞的医学疗法的开发至关重要。为了鉴定分化过程的生物标志物和潜在调节因子，使用含有 16,659 个七十碱基对寡核苷酸的高质量微阵列来比较未分化 hESC 系和分化胚状体的基因表达谱。先前在未分化 hESC 系中鉴定出的“干细胞”基因在分化细胞中表现出下调，而随着细胞分化，一些基因的表达被诱导。此外，194 个基因的子集在人胚状体 (hEB) 中过度表达超过 ≥ 3 倍。其中包括 37 个新基因和 157 个已知基因。基因表达通过多种技术进行验证，包括另一种大规模阵列、逆转录聚合酶链反应、聚焦 cDNA 微阵列、大规模并行特征测序 (MPSS) 分析和免疫细胞化学。几个新的 hEB 特异性表达序列标签 (EST) 被映射到人类基因组数据库并表征了它们的表达谱。层次聚类分析清楚地描绘了未分化和分化 hESC 基因表达谱的明显差异，并证实微阵列分析可以轻松地区分它们。这些结果提供了一组独特基因的详细表征，可用于评估 hESC 分化。
The identification of molecular pathways of differentiation of embryonic stem cells (hESC) is critical for the development of stem cell based medical therapies. In order to identify biomarkers and potential regulators of the process of differentiation, a high quality microarray containing 16,659 seventy base pair oligonucleotides was used to compare gene expression profiles of undifferentiated hESC lines and differentiating embryoid bodies. Previously identified "stemness" genes in undifferentiated hESC lines showed down modulation in differentiated cells while expression of several genes was induced as cells differentiated. In addition, a subset of 194 genes showed overexpression of greater than ≥ 3 folds in human embryoid bodies (hEB). These included 37 novel and 157 known genes. Gene expression was validated by a variety of techniques including another large scale array, reverse transcription polymerase chain reaction, focused cDNA microarrays, massively parallel signature sequencing (MPSS) analysis and immunocytochemisty. Several novel hEB specific expressed sequence tags (ESTs) were mapped to the human genome database and their expression profile characterized. A hierarchical clustering analysis clearly depicted a distinct difference in gene expression profile among undifferentiated and differentiated hESC and confirmed that microarray analysis could readily distinguish them. These results present a detailed characterization of a unique set of genes, which can be used to assess the hESC differentiation.