Skip to main content

Generating Neurons from Stem Cells

  • Protocol
Neural Stem Cells

Summary

Recent work shows that major developmental and clinical processes such as central nervous system regeneration and carcinogenesis involve stem cells (SCs) in the brain. In spite of this importance, the requirements of these SCs and their differentiated offspring (neurons, astrocytes, and oligodendrocytes) for survival and proper function are little understood. In vivo, the SCs themselves interact with their environment. This “SC niche” may be complex because it likely includes cells of the vascular and immune systems. The ability to maintain (1) and differentiate (14) central nervous system (CNS) SCs in tissue culture where they can be pharmacologically or genetically (5) manipulated provides a powerful starting point for understanding their behavior. We present detailed information on the methods that permit CNS SCs to differentiate into functional neurons in tissue culture. Important aspects of the culture systems include (1) homogeneity, so that the input and output of a manipulation is known to involve the SC itself; (2) growth in monolayer to visualize and study individual SCs and their offspring; and (3) the use of fully defined culture components to exclude unknown factors from the culture. These conditions support the differentiation of functional, electrically active neurons. These methods allow cell growth and differentiation from normal adult and diseased tissue derived from both animal models and clinical samples. Ultimate validation of such a system comes from accurate prediction of in vivo effects, and the methods we present for CNS SC culture have also successfully predicted regenerative responses in the injured adult nervous system.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Johe, K. K., Hazel, T. G., Muller, T., Dugich-Djordjevic, M. M., and McKay, R. D. (1996) Single factors direct the differentiation of stem cells from the fetal and adult central nervous system. Genes Dev. 10, 3129–3140.

    Article  CAS  PubMed  Google Scholar 

  2. Rajan, P. and McKay, R. D. (1998) Multiple routes to astrocytic differentiation in the CNS. J. Neurosci. 18, 3620–3629.

    CAS  PubMed  Google Scholar 

  3. Vicario-Abejon, C., Collin, C., Tsoulfas, P., and McKay, R. D. (2000) Hippocampal stem cells differentiate into excitatory and inhibitory neurons. Eur. J. Neurosci. 12, 677–688.

    Article  CAS  PubMed  Google Scholar 

  4. Blondel, O., Collin, C., McCarran, W. J., et al. (2000) A glia-derived signal regulating neuronal differentiation. J. Neurosci. 20, 8012–8020.

    CAS  PubMed  Google Scholar 

  5. Androutsellis-Theotokis, A., Leker, R. R., Soldner, F., et al. (2006) Notch signalling regulates stem cell numbers in vitro and in vivo. Nature 442, 823–826.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the intramural research program at the National Institutes of Neurological Disorders and Stroke.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Androutsellis-Theotokis, A. et al. (2008). Generating Neurons from Stem Cells. In: Weiner, L.P. (eds) Neural Stem Cells. Methods in Molecular Biology™, vol 438. Humana Press. https://doi.org/10.1007/978-1-59745-133-8_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-133-8_4

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-846-1

  • Online ISBN: 978-1-59745-133-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics