Tissue-Specific Stem Cells

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Inhibition of Notch Signaling in Human Embryonic Stem Cell–Derived Neural Stem Cells Delays G1/S Phase Transition and Accelerates Neuronal Differentiation In Vitro and In Vivo†‡§

  1. Lodovica Borghese1,
  2. Dasa Dolezalova2,3,
  3. Thoralf Opitz1,
  4. Simone Haupt1,
  5. Anke Leinhaas1,
  6. Barbara Steinfarz1,
  7. Philipp Koch1,
  8. Frank Edenhofer1,
  9. Ales Hampl2,3,
  10. Oliver Brüstle1,¶,*

Article first published online: 16 MAR 2010

DOI: 10.1002/stem.408

Issue

STEM CELLS

STEM CELLS

Volume 28, Issue 5, pages 955–964, May 2010

Additional Information

How to Cite

Borghese, L., Dolezalova, D., Opitz, T., Haupt, S., Leinhaas, A., Steinfarz, B., Koch, P., Edenhofer, F., Hampl, A. and Brüstle, O. (2010), Inhibition of Notch Signaling in Human Embryonic Stem Cell–Derived Neural Stem Cells Delays G1/S Phase Transition and Accelerates Neuronal Differentiation In Vitro and In Vivo. STEM CELLS, 28: 955–964. doi: 10.1002/stem.408

Author Information

  1. 1

    Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn and Hertie Foundation, Bonn, Germany

  2. 2

    Biology Department, Faculty of Medicine, Masaryk University, Brno, Czech Republic

  3. 3

    Department of Molecular Embryology, Institute of Experimental Medicine, ASCR, v.v.i., Prague, Czech Republic

  1. Telephone: +49-228-6885-500; Fax: +49-228-6885-501

*Institute of Reconstructive Neurobiology, University of Bonn, LIFE & BRAIN Center, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany

  1. †

    Disclosure of potential conflicts of interest is found at the end of this article.

  2. ‡

    Author contributions: L.B.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; D.D.: collection and assembly of data, data analysis and interpretation; T.O.: collection and assembly of data, data analysis and interpretation; S.H.: collection and assembly of data, data analysis and interpretation; A.L.: support with the establishment of in vivo experiments; B.S.: support with the establishment of rat hippocampus slice cultures; P.K.: provision of study material; F.E.: provision of study material, manuscript writing; A.H.: data interpretation, manuscript writing; O.B.: conception and design, data interpretation, manuscript writing, final approval of manuscript.

  3. §

    First published online in STEM CELLS EXPRESS March 16, 2010.

Publication History

  1. Issue published online: 10 MAY 2010
  2. Article first published online: 16 MAR 2010
  3. Accepted manuscript online: 16 MAR 2010 12:00AM EST
  4. Manuscript Accepted: 3 MAR 2010
  5. Manuscript Received: 14 OCT 2009

Funded by

  • EU (LSHG-CT-2006-018739, ESTOOLS; FP7-HEALTH-2007-B-22943-NeuroStemcell)
  • DFG. Grant Numbers: SFB TR3 D2, MSM0021622430, AV0Z50390703, 1M0538
  • Hertie Foundation

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Keywords:

  • Neural stem cells;
  • Notch;
  • Neuron;
  • Cell cycle

Abstract

The controlled in vitro differentiation of human embryonic stem cells (hESCs) and other pluripotent stem cells provides interesting prospects for generating large numbers of human neurons for a variety of biomedical applications. A major bottleneck associated with this approach is the long time required for hESC-derived neural cells to give rise to mature neuronal progeny. In the developing vertebrate nervous system, Notch signaling represents a key regulator of neural stem cell (NSC) maintenance. Here, we set out to explore whether this signaling pathway can be exploited to modulate the differentiation of hESC-derived NSCs (hESNSCs). We assessed the expression of Notch pathway components in hESNSCs and demonstrate that Notch signaling is active under self-renewing culture conditions. Inhibition of Notch activity by the γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) in hESNSCs affects the expression of human homologues of known targets of Notch and of several cell cycle regulators. Furthermore, DAPT-mediated Notch inhibition delays G1/S-phase transition and commits hESNSCs to neurogenesis. Combined with growth factor withdrawal, inhibition of Notch signaling results in a marked acceleration of differentiation, thereby shortening the time required for the generation of electrophysiologically active hESNSC-derived neurons. This effect can be exploited for neural cell transplantation, where transient Notch inhibition before grafting suffices to promote the onset of neuronal differentiation of hESNSCs in the host tissue. Thus, interference with Notch signaling provides a tool for controlling human NSC differentiation both in vitro and in vivo. STEM CELLS 2010;28:955–964

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