Imaging-based chemical screens using normal and glioma-derived neural stem cells
Imaging-based chemical screens using normal and glioma-derived neural stem cells
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DOI：
10.1042/bst0381067
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发表时间：
2010-08-01
影响因子：
3.9
通讯作者：
Pollard, Steven M.
中科院分区：
文献类型：
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作者：
Danovi, Davide;Falk, Anna;Pollard, Steven M.
胚胎干细胞、组织干细胞和肿瘤干细胞最佳培养方法的开发是其在药物筛选中应用的关键基础。我们先前描述了能够将人放射状神经胶质样胎儿NS（神经干）细胞扩增为稳定细胞系的限定的贴壁培养条件。类似的协议证明有效的建立肿瘤起始干细胞系的人脑肿瘤胶质母细胞瘤多形性，我们称之为GNS（神经胶质瘤NS）细胞。其他人最近也衍生出了比NS细胞具有更大的神经元亚型分化潜力的更原始的人NS细胞系，其与早期神经上皮具有相似性，称为内斯（神经上皮干）细胞。在本文中，我们讨论了这些细胞在化学筛选中的实用性，并描述了基于简单高内容实时图像的平台的方法。我们报告了一组160种激酶抑制剂（抑制剂选择I和II; Calbiochem）对内斯细胞的影响，确定了三种ROCK（Rho相关激酶）抑制剂促进内斯细胞培养物的扩增。对于GNS细胞，我们筛选了一组1000种化合物，并证实了我们先前发现的神经递质信号通路调节剂的细胞毒性作用。这些研究为未来更高通量的筛选提供了框架。
The development of optimal culture methods for embryonic, tissue and cancer stem cells is a critical foundation for their application in drug screening. We previously described defined adherent culture conditions that enable expansion of human radial glia-like fetal NS (neural stem) cells as stable cell lines. Similar protocols proved effective in the establishment of tumour-initiating stem cell lines from the human brain tumour glioblastoma multiforme, which we termed GNS (glioma NS) cells. Others have also recently derived more primitive human NS cell lines with greater neuronal subtype differentiation potential than NS cells, which have similarities to the early neuroepithelium, named NES (neuroepithelial stem) cells. In the present paper, we discuss the utility of these cells for chemical screening, and describe methods for a simple high-content live-image-based platform. We report the effects of a panel of 160 kinase inhibitors (Inhibitor Select I and II; Calbiochem) on NES cells, identifying three inhibitors of ROCK (Rho-associated kinase) as promoting the expansion of NES cell cultures. For the GNS cells, we screened a panel of 1000 compounds and confirmed our previous finding of a cytotoxic effect of modulators of neurotransmitter signalling pathways. These studies provide a framework for future higher-throughput screens.