Department of Dental Regenerative Biotechnology, Seoul National University, Seoul, 110-749, Korea.
Mol Cells. 2013 May;35(5):456-61. doi: 10.1007/s10059-013-0083-0. Epub 2013 May 16.
Embryonic stem (ES) cells can be valuable for monitoring differentiation processes and for improving applications in basic developmental biology. The application of ES cells can be a useful tool for drug discovery and toxicology. Therefore, we suggest the high-throughput screening (HTS) system based on ES cells in this study. Firstly, we optimized the feeder-free condition and seeding cell number which can maintained for at least 7 days without over-confluency. We analyzed the system by cell viability, proliferation activity, RT-PCR and morphologic/immunohistochemical evaluations. The optimal cell seeding number was 30/well that was maintained the typical colonial morphology over 9 d with 1,000 U/ml LIF in the limited space. The cell in optimized condition expressed ALP, SSEA-1, Oct 4 and Nanog and the genetic expressions showed similar to protein expressions. The cell lineage marker expressions showed faint or none. The cell viability and proliferation activity were increased in time-dependent manner in our optimized HTS system. In conclusion, the novel HTS system using ES cells can by useful for developing models for drug discovery as well as toxicological screening in the near future.
胚胎干细胞（ES 细胞）可用于监测分化过程，并改进基础发育生物学中的应用。ES 细胞的应用可以成为药物发现和毒理学的有用工具。因此，我们在本研究中建议使用基于 ES 细胞的高通量筛选（HTS）系统。首先，我们优化了无饲养层条件和接种细胞数量，这些条件下细胞至少可维持 7 天而不会过度融合。我们通过细胞活力、增殖活性、RT-PCR 和形态/免疫组织化学评估来分析该系统。最佳的细胞接种数量为 30/孔，在有限的空间中，在 LIF 为 1000U/ml 的条件下，细胞可维持典型的集落形态达 9 天以上。优化条件下的细胞表达碱性磷酸酶（ALP）、SSEA-1、Oct 4 和 Nanog，且基因表达与蛋白表达相似。细胞谱系标记的表达呈微弱或无表达。细胞活力和增殖活性随时间呈依赖性增加。总之，使用 ES 细胞的新型 HTS 系统在不久的将来可能有助于开发药物发现模型和毒理学筛选。
