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Proc Natl Acad Sci U S A. 2017 Feb 14;114(7):E1234-E1242. doi: 10.1073/pnas.1611473114. Epub 2017 Jan 30.

Differentiation of human telencephalic progenitor cells into MSNs by inducible expression of Gsx2 and Ebf1.

Author information

1
Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy; andrea.faedo@unimi.it elena.cattaneo@unimi.it.
2
Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
3
Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy.
4
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy.
5
Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), San Raffaele Scientific Institute, University of Milan Bicocca, 20132 Milan, Italy.
6
Department of Medicine and Surgery, University of Milan Bicocca, 20126 Milan, Italy.
7
Department of Neuroscience, Neuroscience Institute Cavalieri Ottolenghi, 10043 Torino, Italy.
8
Department of Neuroscience, National Institute of Neuroscience, 10043 Torino, Italy.
9
Miltenyi Biotec GmbH, 51429 Bergisch Gladbach, Germany.

Abstract

Medium spiny neurons (MSNs) are a key population in the basal ganglia network, and their degeneration causes a severe neurodegenerative disorder, Huntington's disease. Understanding how ventral neuroepithelial progenitors differentiate into MSNs is critical for regenerative medicine to develop specific differentiation protocols using human pluripotent stem cells. Studies performed in murine models have identified some transcriptional determinants, including GS Homeobox 2 (Gsx2) and Early B-cell factor 1 (Ebf1). Here, we have generated human embryonic stem (hES) cell lines inducible for these transcription factors, with the aims of (i) studying their biological role in human neural progenitors and (ii) incorporating TF conditional expression in a developmental-based protocol for generating MSNs from hES cells. Using this approach, we found that Gsx2 delays cell-cycle exit and reduces Pax6 expression, whereas Ebf1 promotes neuronal differentiation. Moreover, we found that Gsx2 and Ebf1 combined overexpression in hES cells achieves high yields of MSNs, expressing Darpp32 and Ctip2, in vitro as well in vivo after transplantation. We show that hES-derived striatal progenitors can be transplanted in animal models and can differentiate and integrate into the host, extending fibers over a long distance.

KEYWORDS:

Ebf1; Gsx2; HD; MSNs; hES cells

PMID:
28137879
PMCID:
PMC5321017
DOI:
10.1073/pnas.1611473114
[Indexed for MEDLINE]
Free PMC Article

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