Microarray Analysis of Undifferentiated and Differentiated Human Pluripotent Stem Cells
Microarray Analysis of Undifferentiated and Differentiated Human Pluripotent Stem Cells
复制标题
DOI：
10.5772/14354
复制
发表时间：
2011-04
期刊：
影响因子：
--
通讯作者：
Jane Synnergren;P. Sartipy
中科院分区：
文献类型：
--
作者：
Jane Synnergren;P. Sartipy
在过去的十年里，我们对早期人类发育的分子程序的理解取得了巨大的进步。这些进步背后的主要原因可归因于20世纪90年代末成功分离的人类胚胎干细胞（hESC）系。基于其多能性和无限增殖的基本特性，这些独特的细胞为体外研究早期人类发育过程提供了可能。然而，在充分实现这些细胞的潜力之前，还有许多障碍需要克服。一个重要的问题是增加对控制hESCs分化的基因调控机制的理解。各种各样的工具和技术已经被用来操纵和研究hESC的基本特性和功能。此外，对hESCs的功能衍生物的平行分析为其分化为特定细胞系的机制提供了重要的见解。细胞和组织中的全局转录变化可以使用分子技术进行研究，如DNA微阵列、est枚举、MPSS分析和SAGE。这些实验的结果提供了被研究细胞状态的快照。这种方法已被证明非常适合于hESCs“干性”状态的表征，也适用于鉴定参与其分化的关键途径。使用各种hESC细胞系和技术平台建立了大规模的基因表达数据库，随后使用不同的生物信息学方法分析了这些信息。在hESCs中发现了一组高度表达的基因，这些基因被认为参与了保存未分化细胞的多能性和自我更新能力。此外，一些研究集中在表征特异性分化过程的分子特征上。同样，全局表达分析已被证明是一种非常合适的工具，因为在此类实验的背景下可以揭示新的重要机制。最近，分析microRNA （miRNA）的全球表达谱的可能性已经实现，并且专门为检测miRNA物种而设计的基于微阵列的平台现在已经可用。同时分析hESCs及其分化后代的全球mRNA和miRNA表达谱，预计将为未分化和分化状态下细胞中活跃的调控途径提供额外的见解。在本章中，我们将
Over the last decade, a tremendous progress has been made regarding our understanding of the molecular program involved in early human development. The main reason behind these advancements can be ascribed to the successful isolation of human embryonic stem cell (hESC) lines in the late 1990’s. Based on their fundamental properties of pluripotency and unlimited proliferation, these unique cells have provided the possibility to study early human developmental processes in vitro. However, there are many obstacles to overcome before the potential of these cells can be fully realized. One important issue is to increase the understanding about the gene regulatory mechanisms that control the differentiation of hESCs. A wide variety of tools and technologies have been used to manipulate and study basic hESC characteristics and functions. Furthermore, the parallel analysis of functional derivatives of hESCs has provided important insights into the mechanisms that govern their differentiation into specific cell lineages. Global transcriptional changes in cells and tissues can be studied using molecular techniques such as DNA microarray, EST-enumeration, MPSS profiling, and SAGE. The results from such experiments provide a snapshot of the status of the cells under study. This approach has proven well suited for characterization of the “stemness” state of hESCs, but also for the identification of crucial pathways involved in their differentiation. Large scale gene expression databases have been generated using various hESC lines and technical platforms, and subsequently the information has been analyzed using different bioinformatic approaches. A discrete set of genes has been identified which are highly expressed in hESCs, and these genes are considered to be involved in preserving the pluripotency and self-renewal capacity of the undifferentiated cells. Furthermore, several studies have focused on characterizing the molecular signature of specific differentiation processes. Again, global expression analysis has proven to be a very suitable tool since novel important mechanisms can be revealed in the context of such experiments. More recently, the possibility to analyze also the global expression profile of microRNA (miRNA) has been realized, and microarray based platforms designed specifically for the detection of miRNA species are now available. The concurrent analysis of the global mRNA and miRNA expression profiles of hESCs and their differentiated progenies are anticipated to provide additional insights into the regulatory pathways which are active in the cells in the undifferentiated and differentiated states. In the present chapter, we will