title: Phenazopyridine induces and synchronizes neuronal differentiation of embryonic stem cells.
abstract: 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.
authors.affiliation: Department of Pathology and Immunology, University of Geneva Medical School, Switzerland.
authors.forename: David M
authors.initials: DM
authors.lastname: Suter
authors.forename: Olivier
authors.initials: O
authors.lastname: Preynat-Seauve
authors.forename: Diderik
authors.initials: D
authors.lastname: Tirefort
authors.forename: Anis
authors.initials: A
authors.lastname: Feki
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Krause
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D009474:Neurons / Q000378:metabolism*; D010621:Phenazopyridine / Q000494:pharmacology*
chemical_list: D010621:Phenazopyridine