Arrayed cellular microenvironments for identifying culture and differentiation conditions for stem, primary and rare cell populations
Arrayed cellular microenvironments for identifying culture and differentiation conditions for stem, primary and rare cell populations
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DOI：
10.1038/nprot.2012.017
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发表时间：
2012-04-01
期刊：
影响因子：
14.8
通讯作者：
Willert, Karl
中科院分区：
文献类型：
--
作者：
Brafman, David A.;Chien, Shu;Willert, Karl
在生物体的发育过程中，细胞暴露于无数的信号、结构成分和支架中，这些共同构成了细胞微环境。目前的大多数发育生物学研究检查单个或小子集的分子和参数对细胞行为的影响，因此未能探索细胞所暴露的因素的复杂性。在这里，我们描述了一种技术，被称为阵列细胞微环境（ACME），它允许高通量的检查多种细胞外成分的影响，在任何感兴趣的细胞类型的组合方式。我们将特别关注这项技术在人类多能干细胞（hPSC）中的应用，这是一群具有巨大治疗潜力的细胞，其生长和分化条件的特征很差，远未定义和优化。标准的ACME筛选使用先前应用于DNA微阵列的制造和分析的技术，需要标准的细胞培养设施，并且可以在5-10天内从头到尾进行。
During the development of an organism, cells are exposed to a myriad of signals, structural components and scaffolds, which collectively make up the cellular microenvironment. The majority of current developmental biology studies examine the effect of individual or small subsets of molecules and parameters on cellular behavior, and they consequently fail to explore the complexity of factors to which cells are exposed. Here we describe a technology, referred to as arrayed cellular microenvironments (ACMEs), that allows for a high-throughput examination of the effects of multiple extracellular components in a combinatorial manner on any cell type of interest. We will specifically focus on the application of this technology to human pluripotent stem cells (hPSCs), a population of cells with tremendous therapeutic potential, and one for which growth and differentiation conditions are poorly characterized and far from defined and optimized. A standard ACME screen uses the technologies previously applied to the manufacture and analysis of DNA microarrays, requires standard cell-culture facilities and can be performed from beginning to end within 5-10 days.