Department of Pathology and Immunology, University of Geneva Medical School, Switzerland.
J Cell Mol Med. 2009 Sep;13(9B):3517-27. doi: 10.1111/j.1582-4934.2009.00660.x.
Embryonic stem (ES) cells are powerful tools to understand mechanisms of neuronal differentiation and to engineer neurons for in vitro studies and cell therapy. We developed a screening approach to identify small organic molecules driving neuronal differentiation of ES cells. For this purpose, we used a lentivector carrying a dual luciferase reporter system to engineer an ES cell line which allowed us to screen for small organic molecules enhancing neuronal differentiation. One of them, phenazopyridine, was further analysed in human ES cells. Phenazopyridine: (i) enhanced neuronal differentiation, (ii) increased cell survival, (iii) decreased the amount of non-neuronal and undifferentiated cells and (iv) synchronized the cellular differentiation state. Phenazopyridine allowed the development of a differentiation protocol compatible with the generation of clinical grade neural precursors, which were able differentiate into different neuronal subtypes, astrocytes and oligodendrocytes. In summary, we describe a powerful approach to identify small molecules directing stem cell differentiation. This led to the establishment of a new application for an old drug and the development of a novel clinical grade protocol for neuronal differentiation of ES cells.
胚胎干细胞（ES 细胞）是理解神经元分化机制和为体外研究和细胞治疗工程神经元的有力工具。我们开发了一种筛选方法，以鉴定促使 ES 细胞向神经元分化的小分子有机化合物。为此，我们使用携带双荧光素酶报告系统的慢病毒载体来工程化 ES 细胞系，使我们能够筛选增强神经元分化的小分子有机化合物。其中一种名为 phenazopyridine 的化合物在人 ES 细胞中进一步进行了分析。phenazopyridine：（i）增强神经元分化，（ii）增加细胞存活，（iii）减少非神经元和未分化细胞的数量，（iv）使细胞分化状态同步。phenazopyridine 允许开发一种与生成临床级神经前体细胞兼容的分化方案，这些细胞能够分化为不同的神经元亚型、星形胶质细胞和少突胶质细胞。总之，我们描述了一种识别小分子指导干细胞分化的有效方法。这导致一种旧药物的新应用的建立，并开发了一种用于 ES 细胞向神经元分化的新型临床级方案。
