title: Human embryonic stem cell-derived neuronal cells form spontaneously active neuronal networks in vitro.
abstract: The production of functional human embryonic stem cell (hESC)-derived neuronal cells is critical for the application of hESCs in treating neurodegenerative disorders. To study the potential functionality of hESC-derived neurons, we cultured and monitored the development of hESC-derived neuronal networks on microelectrode arrays. Immunocytochemical studies revealed that these networks were positive for the neuronal marker proteins beta-tubulin(III) and microtubule-associated protein 2 (MAP-2). The hESC-derived neuronal networks were spontaneously active and exhibited a multitude of electrical impulse firing patterns. Synchronous bursts of electrical activity similar to those reported for hippocampal neurons and rodent embryonic stem cell-derived neuronal networks were recorded from the differentiated cultures until up to 4 months. The dependence of the observed neuronal network activity on sodium ion channels was examined using tetrodotoxin (TTX). Antagonists for the glutamate receptors NMDA [D(-)-2-amino-5-phosphonopentanoic acid] and AMPA/kainate [6-cyano-7-nitroquinoxaline-2,3-dione], and for GABAA receptors [(-)-bicuculline methiodide] modulated the spontaneous electrical activity, indicating that pharmacologically susceptible neuronal networks with functional synapses had been generated. The findings indicate that hESC-derived neuronal cells can generate spontaneously active networks with synchronous communication in vitro, and are therefore suitable for use in developmental and drug screening studies, as well as for regenerative medicine.
authors.affiliation: Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland.
authors.forename: Teemu J
authors.initials: TJ
authors.lastname: Heikkilä
authors.forename: Laura
authors.initials: L
authors.lastname: Ylä-Outinen
authors.forename: Jarno M A
authors.initials: JM
authors.lastname: Tanskanen
authors.forename: Riikka S
authors.initials: RS
authors.lastname: Lappalainen
authors.forename: Heli
authors.initials: H
authors.lastname: Skottman
authors.forename: Riitta
authors.initials: R
authors.lastname: Suuronen
authors.forename: Jarno E
authors.initials: JE
authors.lastname: Mikkonen
authors.forename: Jari A K
authors.initials: JA
authors.lastname: Hyttinen
authors.forename: Susanna
authors.initials: S
authors.lastname: Narkilahti
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D015374:Biosensing Techniques; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival; D004558:Electric Stimulation / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D018683:Excitatory Amino Acid Agents / Q000494:pharmacology; D018682:GABA Agents / Q000494:pharmacology; D006801:Humans; D008839:Microelectrodes; D009415:Nerve Net / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D018683:Excitatory Amino Acid Agents; D018682:GABA Agents; D026941:Sodium Channel Blockers; D013779:Tetrodotoxin