Expression Profiling Mouse Embryonic/Somatic Stem Cells

小鼠胚胎/体干细胞表达谱分析

• 批准号: 6815285
• 负责人: Minoru S Ko
• 金额: --
• 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/6815285
• 关键词: cell differentiation; developmental genetics; embryonic stem cell; functional /structural genomics; gene expression; gene expression profiling; hematopoietic stem cells; laboratory mouse; mammalian embryology; microarray technology; nerve stem cell; osteoblasts; trophoblast

The goal of this research project is to understand the nature of mouse embryonic and adult stem cells and to identify genes that are responsible for the maintenance of cellular pluripotency. We have been conducting global gene expression profiling with the mouse embryonic cDNA microarrays developed in our laboratory. As a first step, large-scale gene expression profiling was performed on embryo-derived stem cell lines to identify molecular signatures of pluripotency and lineage specificity. Analysis of pluripotent embryonic stem (ES) cells, extraembryonic-restricted trophoblast stem (TS) cells, and terminally-differentiated mouse embryo fibroblast (MEF) cells identified expression profiles unique to each cell type, as well as genes common only to ES and TS cells. Whereas most of the MEF-specific genes had previously been characterized, the majority (67%) of the ES-specific genes was novel and did not include known differentiated cell markers. We suggest that pluripotency requires a set of genes not expressed in other cell types, while lineage-restricted stem cells, like TS cells, express genes predictive of their differentiated lineage. The identification of genes that are specifically expressed in ES cells has provided an important first step for understanding the pluripotency of stem cells. These genes can be used as markers for pluripotent stem cells. Furthermore, the manipulation of these genes in ES cells and other cell types can further elucidate their function and provide possible means to harness the cellular pluripotency. To extend this work and extract the common features of stem cells, we have thus far profiled the following stem cells: (1) Differentiation and lineage commitment of pluripotent mouse embryonic stem (ES) cells. Expression profiling of mouse ES cells at six time points during the course of their differentiation has been performed. (2) Mesenchymal stem cells and derivative osteoblast cells. (3) Neural Stem cells and neuron/glia cells. We are currently working on data analysis and independent validation of results. We also plan to do expression profiling on Embryonic Germ (EG) cells, on ES cells with the altered expression of Stat3, and on ES cells with altered expression of Oct-3/4.

该研究项目的目标是了解小鼠胚胎和成体干细胞的性质，并确定负责维持细胞多能性的基因。我们一直在用实验室开发的小鼠胚胎cDNA微阵列进行全球基因表达谱分析。作为第一步，对胚胎来源的干细胞系进行了大规模的基因表达谱分析，以确定多能性和谱系特异性的分子特征。对多能胚胎干细胞（ES）细胞、胚胎外限制性滋养细胞（TS）细胞和终末分化小鼠胚胎成纤维细胞（MEF）细胞的分析发现了每种细胞类型所特有的表达谱，以及仅在ES和TS细胞中常见的基因。虽然大多数mef特异性基因之前已经被表征，但大多数（67%）es特异性基因是新的，不包括已知的分化细胞标记物。我们认为多能性需要一组在其他细胞类型中不表达的基因，而谱系限制干细胞，如TS细胞，表达预测其分化谱系的基因。胚胎干细胞中特异性表达基因的鉴定为理解干细胞的多能性提供了重要的第一步。这些基因可以作为多能干细胞的标记物。此外，在胚胎干细胞和其他细胞类型中操纵这些基因可以进一步阐明它们的功能，并为利用细胞多能性提供可能的手段。为了扩展这项工作并提取干细胞的共同特征，我们迄今为止概述了以下干细胞：(1)多能小鼠胚胎干细胞（ES）的分化和谱系承诺。对小鼠胚胎干细胞在分化过程中的6个时间点进行了表达谱分析。(2)间充质干细胞和衍生成骨细胞。(3)神经干细胞和神经元/胶质细胞。我们目前正在进行数据分析和结果的独立验证。我们还计划在胚胎胚（EG）细胞、Stat3表达改变的胚胎干细胞和Oct-3/4表达改变的胚胎干细胞上进行表达谱分析。