Expression profiling of mouse embryonic and tissue stem cells

小鼠胚胎和组织干细胞的表达谱

• 批准号: 7964003
• 负责人: Minoru Ko
• 金额: $ 30.06万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7964003
• 关键词: Adult; Aging; Biological Assay; Cell Culture Techniques; Cell Line; Cell Lineage; Cells; DNA Microarray Chip; Development; Ectoderm; Embryo; Endoderm; Fibroblasts; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Germ Cells; Germ Lines; Goals; Half-Life; In Situ Hybridization; In Vitro; Maintenance; Maps; Measures; Messenger RNA; Molecular Profiling; Mus; Organ; Pattern; Population; Positioning Attribute; Principal Component Analysis; Protein Binding; RNA; RNA-Binding Proteins; Replacement Therapy; Research Project Grants; Rest; Spottings; Stem cells; Transcript; Undifferentiated; Work; adult stem cell; cell type; embryo tissue; embryonic stem cell; in vivo; neurodevelopment; pluripotency; relating to nervous system; self-renewal; stem; trophoblast

In our earlier work, we compared the global expression profiles of mouse ES cells and trophoblast stem (TS) cells by DNA microarrays. We studied Esg1, one of the genes identified as a gene expressed specifically in ES cells, and found that the gene encodes an RNA-binding protein that binds to many RNA targets. We have also compared the expression profiles of mouse ES cells undergoing neural differentiation in vitro and those of adult neural stem progenitor (NS) cells. The results suggested that ES cells undergoing neural differentiation in vitro recapitulate the development of neural lineages in vivo. We also inferred a set of 4,000 genes, the expression of which increased with neural commitment differentiation; it can be used as a scale for the degree of commitment to neural differentiation. We also carried out global gene expression profiling of mouse embryonic germ (EG) cells and multipotent adult stem cells (MAPCs). We have carried out high-throughput in situ hybridization assays on ES cell cultures for 244 genes. We found that three genes (Zscan4, Whsc2, and Rhox9) showed a spotty expression pattern (spot-in-colony pattern). We also found nine genes that showed a relatively heterogeneous expression pattern (mosaic-in-colony pattern: Zfp42/Rex1, Rest, Atf4, Pa2g4, E2f2, Nanog, Dppa3/Pgc7/Stella, Esrrb, and Fscn1). This indicates the presence of heterogeneous cell population in undifferentiated ES cell cultures. As the baseline information to understand the gene expression regulation in ES cells, we have measured the mRNA half-life of essentially all mouse genes by using DNA microarrays. Recently, we have also demonstrated that principal component analysis of global gene expression profiles map cells in multidimensional transcript profile space and the positions of differentiating cells progress in a stepwise manner along trajectories starting from undifferentiated embryonic stem (ES) cells located in the apex. We have presented three 'cell lineage trajectories', which represent the differentiation of ES cells into the first three lineages in mammalian development: primitive endoderm, trophoblast and primitive ectoderm/neural ectoderm. The positions of the cells along these trajectories seem to reflect the developmental potency of cells and can be used as a scale for the potential of cells. Indeed, we show that embryonic germ cells and induced pluripotent cells are mapped near the origin of the trajectories, whereas mouse embryo fibroblast and fibroblast cell lines are mapped near the far end of the trajectories.

在我们早期的工作中，我们通过DNA微阵列比较了小鼠胚胎干细胞和滋养细胞（TS）细胞的全局表达谱。我们研究了Esg1，这是在胚胎干细胞中特异性表达的基因之一，发现该基因编码一种RNA结合蛋白，可以结合许多RNA靶点。我们还比较了体外神经分化的小鼠胚胎干细胞和成人神经干细胞（NS）的表达谱。结果表明，体外神经分化的胚胎干细胞在体内可以再现神经谱系的发育。我们还推断出一组4000个基因，其表达随着神经承诺分化而增加；它可以用作神经分化承诺程度的尺度。我们还进行了小鼠胚胎生殖细胞（EG）和多能成体干细胞（MAPCs）的全球基因表达谱分析。我们对胚胎干细胞培养的244个基因进行了高通量原位杂交试验。我们发现三个基因（Zscan4， Whsc2和Rhox9）表现出点状表达模式（点在群体模式）。我们还发现9个基因表现出相对异质的表达模式（嵌合体-集落模式：Zfp42/Rex1、Rest、Atf4、Pa2g4、E2f2、Nanog、Dppa3/Pgc7/Stella、Esrrb和Fscn1）。这表明在未分化的胚胎干细胞培养中存在异质细胞群。