Abstract
The discovery of neural stem cells (NSCs) in the mammalian brain has raised many expectations as these unique cells might recapitulate different neurological diseases, including brain tumors, both from a functional and molecular perspective. Proper in vitro culturing of NSCs has emerged as a critical methodological issue, given that it should preserve the in vivo features of NSCs, with particular emphasis on cell heterogeneity. At the same time, the methodology for NSC culturing should allow the production of large amounts of cells to be exploited not only for prospective clinical applications, but also for drug screening. Direct in vitro selection of NSCs and, very recently, cancer stem cells (CSCs) by means of defined serum-free conditions represents the most reliable methodology to obtain long-term expanding SC lines. Here we describe the methods currently employed to enrich for NSCs/CSCs based on the NeuroSphere Assay (NSA) and their adaptation to specific assays for testing the efficacy of neuroactive compounds.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Winquist RJ, Furey BF, Boucher DM (2010) Cancer stem cells as the relevant biomass for drug discovery. Curr Opin Pharmacol 10:385–390
Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1707–1710
Reynolds BA, Rietze RL (2005) Neural stem cells and neurospheres—re-evaluating the relationship. Nat Methods 2:333–336
Conti L, Pollard SM, Gorba T et al (2005) Niche-independent symmetrical self-renewal of a mammalian tissue stem cell. PLoS Biol 3:e283
Galli R, Binda E, Orfanelli U et al (2004) Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res 64:7011–7021
Pollard SM, Yoshikawa K, Clarke ID et al (2009) Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell 4:568–580
Ponti D, Costa A, Zaffaroni N et al (2005) Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res 65:5506–5511
O’Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445:106–110
Wang S (2009) Anchorage-independent growth of prostate cancer stem cells. Methods Mol Biol 568:151–160
Gritti A, Cova L, Parati EA et al (1995) Basic fibroblast growth factor supports the proliferation of epidermal growth factor-generated neuronal precursor cells of the adult mouse CNS. Neurosci Lett 185:151–154
Reynolds BA, 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
Gritti A, Frolichsthal-Schoeller P, Galli R et al (1999) Epidermal and fibroblast growth factors behave as mitogenic regulators for a single multipotent stem cell-like population from the subventricular region of the adult mouse forebrain. J Neurosci 19:3287–3297
Vescovi AL, Reynolds BA, Fraser DD et al (1993) bFGF regulates the proliferative fate of unipotent (neuronal) and bipotent (neuronal/astroglial) EGF-generated CNS progenitor cells. Neuron 11:951–966
Parras CM, Galli R, Britz O et al (2004) Mash1 specifies neurons and oligodendrocytes in the postnatal brain. EMBO J 23:4495–4505
Diamandis P, Wildenhain J, Clarke ID et al (2007) Chemical genetics reveals a complex functional ground state of neural stem cells. Nat Chem Biol 3:268–273
Danovi D, Falk A, Humphreys P et al (2010) Imaging-based chemical screens using normal and glioma-derived neural stem cells. Biochem Soc Trans 38:1067–1071
Mazzoleni S, Politi LS, Pala M et al (2010) Epidermal growth factor receptor expression identifies functionally and molecularly distinct tumor-initiating cells in human glioblastoma multiforme and is required for gliomagenesis. Cancer Res 70:7500–7513
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 SpringerScience+Business Media New York
About this protocol
Cite this protocol
Galli, R. (2013). The Neurosphere Assay Applied to Neural Stem Cells and Cancer Stem Cells. In: Moll, J., Colombo, R. (eds) Target Identification and Validation in Drug Discovery. Methods in Molecular Biology, vol 986. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-311-4_17
Download citation
DOI: https://doi.org/10.1007/978-1-62703-311-4_17
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-310-7
Online ISBN: 978-1-62703-311-4
eBook Packages: Springer Protocols