Abstract
Available methods for differentiating human embryonic stem cells (ESCs) and induced pluripotent cells (iPSCs) into neurons are often cumbersome, slow, and variable. Alternatively, human fibroblasts can be directly converted into induced neuronal (iN) cells. However, with present techniques conversion is inefficient, synapse formation is limited, and only small amounts of neurons can be generated. Here, we show that human ESCs and iPSCs can be converted into functional iN cells with nearly 100% yield and purity in less than 2 weeks by forced expression of a single transcription factor. The resulting ES-iN or iPS-iN cells exhibit quantitatively reproducible properties independent of the cell line of origin, form mature pre- and postsynaptic specializations, and integrate into existing synaptic networks when transplanted into mouse brain. As illustrated by selected examples, our approach enables large-scale studies of human neurons for questions such as analyses of human diseases, examination of human-specific genes, and drug screening.
Copyright © 2013 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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6-Cyano-7-nitroquinoxaline-2,3-dione / pharmacology
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Animals
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Biophysical Phenomena / genetics
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Biophysical Phenomena / physiology*
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Biophysics
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Brain / cytology
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Calcium / metabolism
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Cells, Cultured
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Collagen Type VII / genetics
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Electric Stimulation
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Epidermolysis Bullosa Dystrophica / genetics
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Epidermolysis Bullosa Dystrophica / pathology
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Excitatory Amino Acid Antagonists / pharmacology
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Excitatory Postsynaptic Potentials / drug effects
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Excitatory Postsynaptic Potentials / genetics
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Fibroblasts
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Gene Expression Regulation / drug effects
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Gene Expression Regulation / genetics
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Gene Expression Regulation / physiology*
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Green Fluorescent Proteins / genetics
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Humans
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Mice
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Microscopy, Confocal
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Munc18 Proteins / genetics
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Munc18 Proteins / metabolism
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Mutation / genetics
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism*
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Neurons / physiology*
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Patch-Clamp Techniques
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Pluripotent Stem Cells / physiology*
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RNA, Small Interfering / physiology
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Rhodopsin / genetics
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Sodium Channel Blockers / pharmacology
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Synapses / physiology
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Tetrodotoxin / pharmacology
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Time Factors
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Transfection
Substances
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COL7A1 protein, human
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Collagen Type VII
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Excitatory Amino Acid Antagonists
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Munc18 Proteins
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Nerve Tissue Proteins
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RNA, Small Interfering
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Sodium Channel Blockers
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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Tetrodotoxin
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6-Cyano-7-nitroquinoxaline-2,3-dione
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Rhodopsin
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Calcium