Small-molecule fluorophores to detect cell-state switching in the context of high-throughput screening.
Small-molecule fluorophores to detect cell-state switching in the context of high-throughput screening.
复制标题
DOI：
10.1021/ja077656d
复制
发表时间：
2008-03
影响因子：
15
通讯作者：
B. Wagner;Hyman A. Carrinski;Y. Ahn;Y. Kim;Tamara J. Gilbert;Dina A. Fomina;S. Schreiber;Young‐Tae Chan
中科院分区：
文献类型：
--
作者：
B. Wagner;Hyman A. Carrinski;Y. Ahn;Y. Kim;Tamara J. Gilbert;Dina A. Fomina;S. Schreiber;Young‐Tae Chan
一种能够区分细胞分化所导致的不同状态的小分子将具有巨大的价值，例如，在再生医学方面的努力。我们筛选了一组荧光小分子，以区分小鼠骨骼肌细胞系的分化状态。C2 C12成肌细胞和肌管的基于荧光的高通量筛选导致鉴定出具有所需选择性的六种化合物，这通过在相同细胞状态下的高含量筛选来证实。将导致细胞状态之间最大荧光强度差异的化合物用作84种激酶抑制剂的中试筛选中的筛选剂，每种激酶抑制剂以四种剂量存在，用于抑制肌生成。在激酶抑制剂中，17种在一种或多种浓度下导致荧光减少;在“命中”中包括已知的肌生成抑制剂，证实该化合物能够检测分化的肌管状态。我们建议，在这里报告的筛选剂筛选的战略可能会在未来更广泛地扩展。
A small molecule capable of distinguishing the distinct states resulting from cellular differentiation would be of enormous value, for example, in efforts aimed at regenerative medicine. We screened a collection of fluorescent small molecules for the ability to distinguish the differentiated state of a mouse skeletal muscle cell line. High-throughput fluorescence-based screening of C2C12 myoblasts and myotubes resulted in the identification of six compounds with the desired selectivity, which was confirmed by high-content screening in the same cell states. The compound that resulted in the greatest fluorescence intensity difference between the cell states was used as the screening agent in a pilot screen of 84 kinase inhibitors, each present in four doses, for inhibition of myogenesis. Of the kinase inhibitors, 17 resulted in reduction of fluorescence at one or more concentrations; among the "hits" included known inhibitors of myogenesis, confirming that this compound is capable of detecting the differentiated myotube state. We suggest that the strategy of screening for screening agents reported here may be extended more broadly in the future.