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Induction of a midbrain dopaminergic phenotype in Nurr1-overexpressing neural stem cells by type 1 astrocytes

Nature Biotechnology volume 17, pages 653–659 (1999)Cite this article

Abstract

The implementation of neural stem cell lines as a source material for brain tissue transplants is currently limited by the ability to induce specific neurochemical phenotypes in these cells. Here, we show that coordinated induction of a ventral mesencephalic dopaminergic phenotype in an immortalized multipotent neural stem cell line can be achieved in vitro. This process requires both the overexpression of the nuclear receptor Nurr1 and factors derived from local type 1 astrocytes. Over 80% of cells obtained by this method demonstrate a phenotype indistinguishable from that of endogenous dopaminergic neurons. Moreover, this procedure yields an unlimited number of cells that can engraft in vivo and that may constitute a useful source material for neuronal replacement in Parkinson's disease.

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Figure 1: Nurr1 overexpression induces the differentiation of C17.2 neural stem cells into nondopaminergic neurons.
Figure 2: The VM is the source of a signal required for the induction of TH-positive neurons from Nurr1-overexpressing neural stem cells.
Figure 3: High proliferative rates in Nurr1-overexpressing clones correlate with high induction of TH-positive neurons from C17.2-Nurr1 in VM co-cultures.
Figure 4: Retinoids and bFGF increase the number of TH-positive neurons derived from C17.2-Nurr1, but not from C17.2 cells, in VM co-cultures.
Figure 5: VM type 1 astrocytes specifically induce a dopaminergic phenotype on C17.2-Nurr1 neural stem cells in SFM.
Figure 6: Early activity of Nurr1 produces long-lasting changes in gene expression in C17.2 cells.
Figure 7: Characterization and long-term stability of the dopaminergic phenotype acquired by Nurr1-c42 cells after differentiation.

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Acknowledgements

We thank Drs. P. Ernfors and B.-A. Sieber for critical reading of the manuscript and discussion, Drs. S. Ferré and M. Höistad for assistance on HPLC, Dr. L. Foley for the generous gift of SR11237, Lotta Johansson for secretarial help, and Annika Ahlsen for additional assistance. Financial support was obtained from the European Commission, Swedish MRC, Karolinska Institute, and the Jeanssonska and Kapten Arthur Eriksson Foundations. E.A. and J.W. were supported by the Karolinska Institute. D.S.C. was supported by a fellowship from the Gulbenkian Foundation (PGDBM) and Programa Praxis XXI. E.Y.S. was supported by grants from the National Institute of Neurological Diseases and Stroke. J.M.C. was supported by a fellowship from the EMBO. Requests for parental C17.2 cells should be addressed to E.Y.S. (e-mail: snyder@A1.TCH.harvard.edu). Correspondence and requests for materials should be addressed to E.A. (e-mail: ernest@cajal.mbb.ki.se).

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Author notes

  1. Josep M. Canals

    Present address: Departament de Biologia Cellular i Anatomia Patològica, Facultat de Medicina, Universitat de Barcelona, IDIBAPS, Casanova 143, 08036, Barcelona , Spain

  2. Peter Åkerud and Diogo S. Castro: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Biochemistry and Biophysics, Laboratory of Molecular Neurobiology, Karolinska Institute, Stockholm, S-17177 , Sweden

    Joseph Wagner, Peter Åkerud, Pontus C. Holm, Josep M. Canals & Ernest Arenas

  2. The Ludwig Institute for Cancer Research, Stockholm Branch, Karolinska Institute, PO Box 240, Stockholm , S-17177, Sweden

    Diogo S. Castro & Thomas Perlmann

  3. Departments of Neurology and Pediatrics, Harvard Medical School and Division of Neuroscience, Children's Hospital, 320 Longwood Ave., Boston, 02115, MA

    Evan Y. Snyder

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Correspondence to Ernest Arenas.

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Wagner, J., Åkerud, P., Castro, D. et al. Induction of a midbrain dopaminergic phenotype in Nurr1-overexpressing neural stem cells by type 1 astrocytes. Nat Biotechnol 17, 653–659 (1999). https://doi.org/10.1038/10862

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