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PLoS One. 2014 Jul 29;9(7):e103418. doi: 10.1371/journal.pone.0103418. eCollection 2014.

A time course analysis of the electrophysiological properties of neurons differentiated from human induced pluripotent stem cells (iPSCs).

Author information

1
Department of Pathology & Cell Biology, Columbia University, New York, New York, United States of America; The Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University, New York, New York, United States of America; Columbia Stem Cell Initiative, Columbia University, New York, New York, United States of America.
2
The New York Stem Cell Foundation Research Institute, New York, New York, United States of America.
3
Department of Pathology & Cell Biology, Columbia University, New York, New York, United States of America; The Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University, New York, New York, United States of America; Columbia Stem Cell Initiative, Columbia University, New York, New York, United States of America; Research Center for Neuroscience, Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhonpathom, Thailand.
4
Department of Pathology & Cell Biology, Columbia University, New York, New York, United States of America; Department of Neurology, Columbia University, New York, New York, United States of America.

Abstract

Many protocols have been designed to differentiate human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) into neurons. Despite the relevance of electrophysiological properties for proper neuronal function, little is known about the evolution over time of important neuronal electrophysiological parameters in iPSC-derived neurons. Yet, understanding the development of basic electrophysiological characteristics of iPSC-derived neurons is critical for evaluating their usefulness in basic and translational research. Therefore, we analyzed the basic electrophysiological parameters of forebrain neurons differentiated from human iPSCs, from day 31 to day 55 after the initiation of neuronal differentiation. We assayed the developmental progression of various properties, including resting membrane potential, action potential, sodium and potassium channel currents, somatic calcium transients and synaptic activity. During the maturation of iPSC-derived neurons, the resting membrane potential became more negative, the expression of voltage-gated sodium channels increased, the membrane became capable of generating action potentials following adequate depolarization and, at day 48-55, 50% of the cells were capable of firing action potentials in response to a prolonged depolarizing current step, of which 30% produced multiple action potentials. The percentage of cells exhibiting miniature excitatory post-synaptic currents increased over time with a significant increase in their frequency and amplitude. These changes were associated with an increase of Ca2+ transient frequency. Co-culturing iPSC-derived neurons with mouse glial cells enhanced the development of electrophysiological parameters as compared to pure iPSC-derived neuronal cultures. This study demonstrates the importance of properly evaluating the electrophysiological status of the newly generated neurons when using stem cell technology, as electrophysiological properties of iPSC-derived neurons mature over time.

PMID:
25072157
PMCID:
PMC4114788
DOI:
10.1371/journal.pone.0103418
[Indexed for MEDLINE]
Free PMC Article
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