Abstract
Since the discovery of neural stem cells (NSC) in the embryonic and adult mammalian central nervous system (CNS), there have been a growing numbers of tissue culture media and protocols to study and functionally characterize NSCs and its progeny in vitro. One of these culture systems introduced in 1992 is referred to as the Neurosphere Assay, and it has been widely used to isolate, expand, differentiate and even quantify NSC populations. Several years later because its application as a quantitative in vitro assay for measuring NSC frequency was limited, a new single-step semisolid based assay, the Neural Colony Forming Cell (NCFC) assay was developed to accurately measure NSC numbers. The NCFC assay allows the discrimination between NSCs and progenitors by the size of colonies they produce (i.e., their proliferative potential). The evolution and continued improvements made to these tissue culture tools will facilitate further advances in the promising application of NSCs for therapeutic use.
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References
Reynolds B, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1701–1710
Reynolds BA, Tetzlaff W, Weiss S (1992) A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci 12:4565–4574
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
Morshead CM, Reynolds BA, Craig CG, McBurney MW, Staines WA, Morassutti D, Weiss S, van der Kooy D (1994) Neural stem cells in the adult mammalian forebrain: a relatively quiescent subpopulation of subependymal cells. Neuron 13:1071–1082
Tropepe V, Sibilia M, Ciruna BG, Rossant J, Wagner EF, van der Kooy D (1999) Distinct neural stem cells proliferate in response to EGF and FFG in the developing mouse telencephalon. Dev Biol 208:166–188
Gross CG (2000) Neurogenesis in the adult brain: death of a dogma. Nat Rev Neurosci 1:67–73
Ray J, Peterson DA, Schinstine M, Gage FH (1993) Proliferation, differentiation, and long-term culture of primary hippocampal neurons. Proc Natl Acad Sci USA 90:3602–3606
Conti L, Pollard SM, Gorba T et al (2005) Niche-independent symmetrical self-renewal of a mammalian tissue stem cell. PLoS Biol 3:e283
Conti L, Cattaneo E (2010) Neural stem cell systems: physiological players or in vitro entities? Nat Rev Neurosci 11:176–187
O’Connor TJ, Vescovi AL, Reynolds BA (1998) Isolation and propagation of stem cells from various regions of the embryonic mammalian central nervous system. In: Celis JE (ed) Cell biology: a laboratory handbook, 2nd edn. Academic, New York
Reynolds BA, Rietze RL (2005) Neural stem cells and neurospheres – re-evaluating the relationship. Nat Methods 2:333–336
Rietze RL, Reynolds BA (2006) Neural stem cell isolation and characterization. Methods Enzymol 419:3–23
Singec I, Knoth R, Meyer RP et al (2006) Defining the actual sensitivity and specificity of the neurosphere assay in stem cell biology. Nat Methods 3:801–806
Louis SA, Rietze RL, Deleyrolle L et al (2008) Enumeration of neural stem and progenitor cells in the neural colony-forming cell assay. Stem Cells 26(4):988–996
Acknowledgments
We would like to acknowledge our collaborators Drs. Brent Reynolds, Rod L. Rietze, and Angelo L. Vescovi for their continued technical and scientific help and discussions.
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Louis, S.A., Mak, C.K.H., Reynolds, B.A. (2013). Methods to Culture, Differentiate, and Characterize Neural Stem Cells from the Adult and Embryonic Mouse Central Nervous System. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_30
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DOI: https://doi.org/10.1007/978-1-62703-128-8_30
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