Advertisement

Dopaminergic Neuronal Differentiation Protocol for Human Mesenchymal Stem Cells

  • Katarzyna A. Trzaska
  • Pranela RameshwarEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 698)

Abstract

The generation of dopamine (DA) neurons from stem cells holds great promise for future biomedical research and in the clinical treatment of neurodegenerative diseases, such as Parkinson’s disease. Mesenchymal stem cells (MSCs) derived from the adult human bone marrow (BM) can be easily isolated and expanded in culture while maintaining their immense plasticity. Here, we describe a protocol to generate DA-producing cells from adult human MSCs using a cocktail that includes sonic hedgehog (SHH), fibroblast growth factor 8 (FGF8), and basic fibroblast growth factor (bFGF). Electrophysiological functional DA neurons could be achieved by further treatment with brain-derived neurotrophic factor (BDNF). In summary, a protocol is described for the induction of primary BM-derived human MSCs to specific transdifferentiation; in this case, functional DA neurons. The MSC-derived DA cells express DA-specific markers, synthesize, and secrete dopamine. The described method could be used to generate DA cells for various model systems in which DA-producing cells are implicated in pathophysiological conditions.

Key words

Mesenchymal stem cell Neuron Transdifferentiation Dopamine Sonic hedgehog Fibroblast growth factor Brain-derived growth factor 

Notes

Acknowledgments

This work has been supported by the F.M. Kirby Foundation.

References

  1. 1.
    Picinich, S.C., Mishra, P.J., Mishra, P.J., Glod, J., Banerjee, D. (2007) The therapeutic potential of mesenchymal stem cells. Cell- & tissue-based therapy. Expert Opin Biol Ther 7, 965–73.PubMedCrossRefGoogle Scholar
  2. 2.
    Zipori, D. (2004) Mesenchymal stem cells: harnessing cell plasticity to tissue and organ repair. Blood Cells Mol Dis 33, 211–5.PubMedCrossRefGoogle Scholar
  3. 3.
    Giordano, A., Galderisi, U., Marino, I.R. (2007) From the laboratory bench to the patient’s bedside: an update on clinical trials with mesenchymal stem cells. J Cell Physiol 211, 27–35.PubMedCrossRefGoogle Scholar
  4. 4.
    Phinney, D.G., Isakova, I. (2005) Plasticity and therapeutic potential of mesenchymal stem cells in the nervous system. Curr Pharm Des 11, 1255–65.PubMedCrossRefGoogle Scholar
  5. 5.
    Bianco, P., Riminucci, M., Gronthos, S., Robey, P.G. (2001) Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells 19, 180–92.PubMedCrossRefGoogle Scholar
  6. 6.
    Potian, J.A., Aviv, H., Ponzio, N.M., Harrison, J.S., Rameshwar, P. (2003) Veto-like activity of mesenchymal stem cells: functional discrimination between cellular responses to alloantigens and recall antigens. J Immunol 171, 3426–34.PubMedGoogle Scholar
  7. 7.
    Krampera, M., Pasini, A., Pizzolo, G., Cosmi, L., Romagnani, S., Annunziato, F. (2006) Regenerative and immunomodulatory potential of mesenchymal stem cells. Curr Opin Pharmacol 6, 435–41.PubMedCrossRefGoogle Scholar
  8. 8.
    Greco, S.J., Zhou, C., Ye, J.H., Rameshwar, P. (2007) An interdisciplinary approach and characterization of neuronal cells transdifferentiated from human mesenchymal stem cells. Stem Cells Dev 16, 811–26.PubMedCrossRefGoogle Scholar
  9. 9.
    Tropel, P., Platet, N., Platel, J.C., Noel, D., Albrieux, M., Benabid, A.L., Berger, F. (2006) Functional neuronal differentiation of bone marrow-derived mesenchymal stem cells. Stem Cells 24, 2868–76.PubMedCrossRefGoogle Scholar
  10. 10.
    Sato, Y., Araki, H., Kato, J., Nakamura, K., Kawano, Y., Kobune, M., Sato, T., Miyanishi, K., Takayama, T., Takahashi, M., Takimoto, R., Iyama, S., Matsunaga, T., Ohtani, S., Matsuura, A., Hamada, H., Niitsu, Y. (2005) Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. Blood 106, 756–63.PubMedCrossRefGoogle Scholar
  11. 11.
    Zeng, X., Cai, J., Chen, J., Luo, Y., You, Z.B., Fotter, E., Wang, Y., Harvey, B., Miura, T., Backman, C., Chen, G.J., Rao, M.S., Freed, W.J. (2004) Dopaminergic differentiation of human embryonic stem cells. Stem Cells 22, 925–40.PubMedCrossRefGoogle Scholar
  12. 12.
    Lindvall, O., Kokaia, Z., Martinez-Serrano, A. (2004) Stem cell therapy for human neurodegenerative disorders-how to make it work. Nat Med 10 Suppl, S42–50.PubMedCrossRefGoogle Scholar
  13. 13.
    Correia, A.S., Anisimov, S.V., Li, J.Y., Brundin, P. (2005) Stem cell-based therapy for Parkinson’s disease. Ann Med 37, 487–98.PubMedCrossRefGoogle Scholar
  14. 14.
    Trzaska, K.A., Kuzhikandathil, E.V., Rameshwar, P. (2007) Specification of a dopaminergic phenotype from adult human mesenchymal stem cells. Stem Cells 25, 2797–808.PubMedCrossRefGoogle Scholar
  15. 15.
    Trzaska, K.A., King, C.C., Le, K.Y., Kuzhikandathil, E.V., Nowycky, M.C., Ye, J.H., Rameshwar, P. (2009) Functional maturation of mesenchymal stem cell-derived dopamine progenitors by brain-derived neurotrophic factor. J Neurochem 110, 1058–69.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Medicine Hematology/OncologyUniversity of Medicine and Dentistry of New Jersey – New Jersey Medical SchoolNewarkUSA

Personalised recommendations