Journal
EXPERIMENTAL NEUROLOGY
Volume 218, Issue 1, Pages 109-116Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.expneurol.2009.04.011
Keywords
Human embryonic stem cell-derived neurons; hESC-derived neuronal networks; MEA; Microelectrode array; Neuronal differentiation; Neuronal network; Polyethyleneimine; Spike train; Stem cells; Synchronous bursting
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Funding
- Competitive Research Funding of Pirkanmaa Hospital District, Finland
- Employment and Economic Development Center for Pirkanmaa, Finland
- Arvo and Lea YIppo Foudation, Finland
- Academy of Finland
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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 microelectrocle arrays. Immunocytochemical studies revealed that these networks were positive for the neuronal marker proteins beta-tubulin([I]) 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 GABA(A) 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. (C) 2009 Elsevier Inc. All rights reserved.
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