Summary
During development and continuing into adulthood, stem cells function as a reservoir of undifferentiated cell types, whose role is to support cell genesis in several tissues and organs. In the adult, they play an essential homeostatic role by replacing differentiated cells that are lost due to physiological turnover, injury, or disease. The discovery of such cells in the adult mammalian central nervous system (CNS), an organ traditionally thought to have little or no regenerative capacity, has opened the door to the possibility of designing innovative regenerative therapeutics, an unexpected concept in neurobiology 15 years ago.
In 1992, to detect precursor cells in the adult brain, we employed a serum-free culture system whereby the majority of primary differentiated CNS cells did not survive but a small population of EGF-responsive cells were maintained in an undifferentiated state and proliferated to form clusters, called neurospheres (Reynolds and Weiss, Science 255:1707–1710, 1992). These neurospheres could be (a) dissociated to form numerous secondary spheres or (b) induced to differentiate, generating the three major cell types of the CNS. This chapter outlines the adult mammalian NSC culture methodology and provides technical details of the neurosphere assay to achieve reproducible cultures.
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References
Gross, C. G. (2000). Neurogenesis in the adult brain: death of a dogma. Nat Rev Neurosci 1, 67–73.
Reynolds, B. A., and Weiss, S. (1992). Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255, 1707–10.
Reynolds, B. A., and Weiss, S. (1996). Clonal and population analyses demonstrate that an EGF- responsive mammalian embryonic CNS precursor is a stem cell. Dev Biol 175, 1–13.
Potten, C. S., and Loeffler, M. (1990). Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the crypt. Development 110, 1001–20.
Reynolds, B. A., and Rietze, R. L. (2005). Neural stem cells and neurospheres-re-evaluating the relationship. Nat Methods 2, 333–6.
Evans, G. S., and Potten, C. S. (1991) Stem cells and the elixir of life. Bioessays 13, 135–8.
Foroni, C., Galli, R., Cipelletti, B., Caumo, A., Alberti, S., Fiocco, R., and Vescovi, A. (2007). Resilience to transformation and inherent genetic and functional stability of adult neural stem cells ex vivo. Cancer Res 67, 3725–33.
Hall, P. A., and Watt, F. M. (1989). Stem cells: the generation and maintenance of cellular diversity. Development 106, 619–33.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Deleyrolle, L.P., Reynolds, B.A. (2009). Isolation, Expansion, and Differentiation of Adult Mammalian Neural Stem and Progenitor Cells Using the Neurosphere Assay. In: Gordon, D., Scolding, N. (eds) Neural Cell Transplantation. Methods in Molecular Biology™, vol 549. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-931-4_7
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DOI: https://doi.org/10.1007/978-1-60327-931-4_7
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