Skip to main content

Production and Analysis of Neurospheres from Acutely Dissociated and Postmortem CNS Specimens

  • Protocol
Neural Stem Cells: Methods and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 198))

Abstract

For most of the history of neuroscience, it was widely believed, despite isolated reports to the contrary (1), that de novo generation of neurons in the mammalian CNS did not persist past perinatal development. It was not until the last decade of the twentieth century that the existence, within the CNS of fully developed mammals, of a persistent germinal zone containing neural stem cells (NSCs) with the capacity to differentiate into both neurons and glia became widely accepted (2). It is now known that in vivo genesis of new neurons occurs throughout life, and is restricted primarily to the periventricular subependymal zone (SEZ), which generates neurons destined for the olfactory bulb, and the subgranular layer of the hippocampus, which generates neurons destined for the dentate gyrus (3,4).

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 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Altman, J. (1969) Autoradiographic and histological studies of postnatal neuro-genesis. IV. Cell proliferation and migration in the anterior forebrain, with special reference to persisting neurogenesis in the olfactory bulb. J. Comp. Neurol. 137, 433–458.

    Article  PubMed  CAS  Google Scholar 

  2. Alvarez-Buylla, A. and Lois, C. (1995) Neuronal stem cells in the brain of adult vertebrates. Stem Cells 13, 263–272.

    Article  PubMed  CAS  Google Scholar 

  3. Gage, F., Ray, J. and Fisher, L.(1995) Isolation, characterization, and use of stem cells from the CNS. Ann. Rev. Neurosci. 18, 159–192.

    Article  PubMed  CAS  Google Scholar 

  4. McKay, R. (1997) Stem cells in the central nervous system. Science 276, 66–71.

    Article  PubMed  CAS  Google Scholar 

  5. Kukekov, V. G., Laywell, E. D., Suslov, O. N., Thomas, L. B, Scheffler, B., Davies, K., O’Brien, T. F., Kusakabe, M., and Steindler, D. A. (1999) Multipotent stem/progenitor cells with similar properties arise from two neurogenic regions of adult human brain. Exp. Neurol. 156, 333–344.

    Article  PubMed  CAS  Google Scholar 

  6. Palmer, T. D., Markakis, E. A., Willhoite, A. R., Safar, R., and Gage, E H. (1999) Fibroblast growth factor-2 activates a latent neurogenic program in neural stem cells from diverse regions of the adult CNS. J. Neurosci. 109, 8487–8497.

    Google Scholar 

  7. Scheffler, B., Horn, M., Bluemcke, I., Kukekov, V., Laywell, E. D., and Steindler, D. A. (1999) Marrow-mindedness: a perspective on neuropoiesis. Trends. Neurosci. 22, 348–357.

    Article  PubMed  CAS  Google Scholar 

  8. Richards, L. J., Kilpatrick, T. J., and Bartlett, P. E (1992) De novo generation of neuronal cells from the adult mouse brain. Proc. Natl. Acad. Sci. USA 89, 8591–8595.

    Article  PubMed  CAS  Google Scholar 

  9. Reynolds, B. A. and Weiss, S. (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255, 1707–1710.

    Article  PubMed  CAS  Google Scholar 

  10. Reynolds, B. A., Tetzlaff, W., and Weiss, S. (1992) A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J. Neurosci. 12, 4565–4574.

    PubMed  CAS  Google Scholar 

  11. Kukekov, V. G., Laywell, E. D., Thomas, L. B., and Steindler, D. A. (1997) A nestin-negative precursor cell from the adult mouse brain gives rise to neurons and glia. GLIA 21, 399–407.

    Article  PubMed  CAS  Google Scholar 

  12. Laywell, E. D., Kukekov, V. G., and Steindler, D. A. (1999) Multipotent neuro-spheres can be derived from forebrain subependymal zone and spinal cord of adult mice after protracted postmortem intervals. Exp. Neurol. 156, 430–433.

    Article  PubMed  CAS  Google Scholar 

  13. Laywell, E. D., Rakic, P., Kukekov, V. G., Holland, E. C., and Steindler, D. A. (2000) Identification of a multipotent astrocytic stem cell in the immature and adult mouse brain. Proc. Natl. Acad. Sci. USA 97, 13,883–13,888.

    Article  PubMed  CAS  Google Scholar 

  14. Suslov, O. N., Kukekov, V. G., Laywell, E. D., Scheffler, B., and Steindler, D. A. (2000) RT-PCR amplification of mRNA from single brain neurospheres.J. Neurosci. Meth. 96, 57–61.

    Article  CAS  Google Scholar 

  15. Doetsch, F., Caille, I., Lim, D. A., Garcia-Verdugo, J. M., and Alvarez-Buylla, A. (1999) Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell 97, 703–716.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Lay well, E.D., Kukekov, V.G., Suslov, O., Zheng, T., Steindler, D.A. (2002). Production and Analysis of Neurospheres from Acutely Dissociated and Postmortem CNS Specimens. In: Zigova, T., Sanberg, P.R., Sanchez-Ramos, J.R. (eds) Neural Stem Cells: Methods and Protocols. Methods in Molecular Biology™, vol 198. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-186-8:015

Download citation

  • DOI: https://doi.org/10.1385/1-59259-186-8:015

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-964-3

  • Online ISBN: 978-1-59259-186-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics