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Nat Methods. 2017 Jun;14(6):621-628. doi: 10.1038/nmeth.4291. Epub 2017 May 15.

Generation of pure GABAergic neurons by transcription factor programming.

Author information

1
Institute for Stem Cell Biology and Regenerative Medicine and Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
2
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California, USA.
3
Department of Neurological Surgery and the Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, California, USA.
4
Program in Epithelial Biology and Center for Personal Dynamic Regulomes, Stanford University School of Medicine, Stanford, California, USA.
5
Institute for Computational and Mathematical Engineering, Stanford University School of Medicine, Stanford, California, USA.
6
Department of Psychiatry, University of California, San Francisco, San Francisco, California, USA.

Abstract

Approaches to differentiating pluripotent stem cells (PSCs) into neurons currently face two major challenges-(i) generated cells are immature, with limited functional properties; and (ii) cultures exhibit heterogeneous neuronal subtypes and maturation stages. Using lineage-determining transcription factors, we previously developed a single-step method to generate glutamatergic neurons from human PSCs. Here, we show that transient expression of the transcription factors Ascl1 and Dlx2 (AD) induces the generation of exclusively GABAergic neurons from human PSCs with a high degree of synaptic maturation. These AD-induced neuronal (iN) cells represent largely nonoverlapping populations of GABAergic neurons that express various subtype-specific markers. We further used AD-iN cells to establish that human collybistin, the loss of gene function of which causes severe encephalopathy, is required for inhibitory synaptic function. The generation of defined populations of functionally mature human GABAergic neurons represents an important step toward enabling the study of diseases affecting inhibitory synaptic transmission.

PMID:
28504679
PMCID:
PMC5567689
DOI:
10.1038/nmeth.4291
[Indexed for MEDLINE]
Free PMC Article

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