title: Embryonic stem cells express neuronal properties in vitro.
abstract: Mouse embryonic stem (ES) cells cultured as aggregates and exposed to retinoic acid are induced to express multiple phenotypes normally associated with neurons. A large percentage of treated aggregates produce a rich neuritic outgrowth. Dissociating the induced aggregates with trypsin and plating the cells as a monolayer results in cultures in which a sizable percentage of the cells have a neuronal appearance. These neuron-like cells express class III beta-tubulin and the neurofilament M subunit. Induced cultures express transcripts for neural-associated genes including the neurofilament L subunit, glutamate receptor subunits, the transcription factor Brn-3, and GFAP. Levels of neurofilament L and GAD67 and GAD65 transcripts rise dramatically upon induction. Physiological studies show that the neuron-like cells generate action potentials and express TTX-sensitive sodium channels, as well as voltage-gated potassium channels and calcium channels. We conclude that a complex system of neuronal gene expression can be activated in cultured ES cells. This system should be favorable for investigating some of the mechanisms that regulate neuronal differentiation.
authors.affiliation: Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: G
authors.initials: G
authors.lastname: Bain
authors.forename: D
authors.initials: D
authors.lastname: Kitchens
authors.forename: M
authors.initials: M
authors.lastname: Yao
authors.forename: J E
authors.initials: JE
authors.lastname: Huettner
authors.forename: D I
authors.initials: DI
authors.lastname: Gottlieb
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D018076:DNA, Complementary; D004622:Embryo, Mammalian / Q000166:cytology; D008564:Membrane Potentials; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D014212:Tretinoin / Q000494:pharmacology*; D014404:Tubulin / Q000096:biosynthesis
chemical_list: D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D014404:Tubulin; D014212:Tretinoin