Abstract
Neural stem cells are defined as cells that either gives rise to or derives from the cells of the central nervous system and have the unique properties of stem cells, i.e. self-renewal and multipotentiality. One of the widely used methods of expanding neural stem cells under culture conditions is based on the capacity of these cells to divide continuously when cultured in serum-free medium supplemented with various growth factors. One common method used is to grow neural stem cells as free-floating aggregates of cells called neurospheres. Neurospheres can be generated from several structures of the embryonic and adult mammalian brain. Although viable lines can be generated from crude extracts of brain, a precise dissection is crucial to get a pure population of cells. Here we describe methods for dissection, isolation and generation of neurospheres from embryonic ganglionic eminences and adult subventricular zone of mice and rats.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Reynolds, B. A., and Weiss, S. (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 27, 1707–1710.
Campos, L. S., Leone, D. P., Relvas, J. B., Brakebusch, C., Fassler, R., Suter, U., and French-Constant, C. (2004) Beta1 integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance. Development 14, 3433–3444.
Dunnett, S. B., and Björklund, B. (1992) Staging and dissection of rat embryos in Neural transplantation a practical approach, Dunnett, S. B., and Björklund, B. (Eds.). Oxford University Press, Oxford, pp. 1–18.
Johansson, C. B., Momma, S., Clarke, D. L., Risling, M., Lendahl, U., and Frisen, J. (1999) Identification of a neural stem cell in the adult mammalian central nervous system. Cell 1, 25–34.
Wachs, F. P., Couillard-Despres, S., Engelhardt, M., Wilhelm, D., Ploetz, S., Vroemen, M., Kaesbauer, J., Uyanik, G., Klucken, J., Karl, C., Tebbing, J., Svendsen, C., Weidner, N., Kuhn, H. G., Winkler, J., and Aigner, L. (2003) High efficacy of clonal growth and expansion of adult neural stem cells. Lab. Invest. 7, 949–962.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ahlenius, H., Kokaia, Z. (2010). Isolation and Generation of Neurosphere Cultures from Embryonic and Adult Mouse Brain. In: Ward, A., Tosh, D. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 633. Humana Press. https://doi.org/10.1007/978-1-59745-019-5_18
Download citation
DOI: https://doi.org/10.1007/978-1-59745-019-5_18
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-772-3
Online ISBN: 978-1-59745-019-5
eBook Packages: Springer Protocols