Kunisada Yuya, Shoji Masanobu, Hosoya Masaki
Biology Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.
J Biomol Screen. 2012 Feb;17(2):140-51. doi: 10.1177/1087057111422101. Epub 2011 Sep 30.
Low molecular weight compounds have been shown to be useful for controlling stem cells, and various high-throughput screening systems have been developed for identifying compounds that regulate the differentiation of stem cells. However, the effects of such compounds on stem cell differentiation are usually evaluated by assessing a single parameter, which is insufficient for proper monitoring of the cellular status. In this study, to classify a number of compounds, the authors established a gene expression-based screening system using mouse embryonic stem (ES) cells that monitored multiple parameters. ES cells were differentiated into three germ layers by embryoid body formation and then treated with the test compounds. Next, cellular changes were assessed by analyzing the expression of multiple genes with the multiplex quantitative reverse transcriptase polymerase chain reaction. By screening a library of pharmacologically active compounds with this system, the authors were able to classify 52 compounds that influenced the gene expression profile of ES cells. They also found that some compounds identified by screening could enhance osteogenic or adipogenic differentiation of human mesenchymal stem cells. These results indicate that the screening system is effective for identifying compounds involved in regulating the differentiation of both ES cells and adult stem cells.
低分子量化合物已被证明可用于控制干细胞，并且已经开发了各种高通量筛选系统来鉴定调节干细胞分化的化合物。然而，此类化合物对干细胞分化的影响通常通过评估单个参数来评估，这对于正确监测细胞状态是不够的。在本研究中，为了对多种化合物进行分类，作者使用监测多个参数的小鼠胚胎干细胞建立了基于基因表达的筛选系统。通过胚状体形成将胚胎干细胞分化为三个胚层，然后用测试化合物处理。接下来，通过多重定量逆转录聚合酶链反应分析多个基因的表达来评估细胞变化。通过用该系统筛选药理活性化合物文库，作者能够对52种影响胚胎干细胞基因表达谱的化合物进行分类。他们还发现，通过筛选鉴定出的一些化合物可以增强人间充质干细胞的成骨或成脂分化。这些结果表明，该筛选系统对于鉴定参与调节胚胎干细胞和成人干细胞分化的化合物是有效的。
