Abstract
Neural precursor cells (NPCs) are a renewable cell source that can proliferate and expand for long periods of time and give rise to the main neural cell types of the central nervous system (CNS). Establishing simple and reproducible growth culture conditions is of great importance to study the biology of NPCs and to understand the molecular basis of their behavior in healthy and diseased conditions.
Here, we describe a simple free-floating , serum-free culture condition, known as the neurosphere assay, which is the most commonly used method for the isolation and expansion of NPCs harvested from the adult and fetal CNS. This culture system will result in large numbers of undifferentiated NPC progenies that represents a useful cell source for many in vitro and in vivo applications.
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References
Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255(5052):1707–1710. https://doi.org/10.1126/science.1553558
Reynolds BA, Tetzlaff W, Weiss S (1992) A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci 12(11):4565–4574
Gross CG (2000) Neurogenesis in the adult brain: death of a dogma. Nat Rev Neurosci 1(1):67–73. https://doi.org/10.1038/35036235
Reynolds BA, Rietze RL (2005) Neural stem cells and neurospheres—re-evaluating the relationship. Nat Methods 2(5):333–336. https://doi.org/10.1038/nmeth758
Louis SA, Rietze RL, Deleyrolle L, Wagey RE, Thomas TE, Eaves AC, Reynolds BA (2008) Enumeration of neural stem and progenitor cells in the neural colony-forming cell assay. Stem Cells 26(4):988–996. https://doi.org/10.1634/stemcells.2007-0867
Bez A, Corsini E, Curti D, Biggiogera M, Colombo A, Nicosia RF, Pagano SF, Parati EA (2003) Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Res 993(1–2):18–29. https://doi.org/10.1016/j.brainres.2003.08.061
Azari H, Louis SA, Sharififar S, Vedam-Mai V, Reynolds BA (2011) Neural-colony forming cell assay: an assay to discriminate bona fide neural stem cells from neural progenitor cells. J Vis Exp (49). https://doi.org/10.3791/2639
Pastrana E, Silva-Vargas V, Doetsch F (2011) Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell Stem Cell 8(5):486–498. https://doi.org/10.1016/j.stem.2011.04.007
Azari H, Reynolds BA (2016) In vitro models for neurogenesis. Cold Spring Harb Perspect Biol 8(6). https://doi.org/10.1101/cshperspect.a021279
Azari H, Sharififar S, Rahman M, Ansari S, Reynolds BA (2011) Establishing embryonic mouse neural stem cell culture using the neurosphere assay. J Vis Exp (47). https://doi.org/10.3791/2457
Azari H, Rahman M, Sharififar S, Reynolds BA (2010) Isolation and expansion of the adult mouse neural stem cells using the neurosphere assay. J Vis Exp (45). https://doi.org/10.3791/2393
Aligholi H, Hassanzadeh G, Gorji A, Azari H (2016) A novel biopsy method for isolating neural stem cells from the subventricular zone of the adult rat brain for autologous transplantation in CNS injuries. Methods Mol Biol 1462:711–731. https://doi.org/10.1007/978-1-4939-3816-2_39
Webb RL, Kaiser EE, Jurgielewicz BJ, Spellicy S, Scoville SL, Thompson TA, Swetenburg RL, Hess DC, West FD, Stice SL (2018) Human neural stem cell extracellular vesicles improve recovery in a porcine model of ischemic stroke. Stroke 49(5):1248–1256. https://doi.org/10.1161/STROKEAHA.117.020353
Rong Y, Liu W, Wang J, Fan J, Luo Y, Li L, Kong F, Chen J, Tang P, Cai W (2019) Neural stem cell-derived small extracellular vesicles attenuate apoptosis and neuroinflammation after traumatic spinal cord injury by activating autophagy. Cell Death Dis 10(5):340. https://doi.org/10.1038/s41419-019-1571-8
Rietze RL, Reynolds BA (2006) Neural stem cell isolation and characterization. Methods Enzymol 419:3–23. https://doi.org/10.1016/S0076-6879(06)19001-1
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This work was supported by the Overstreet Foundation.
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Azari, H. (2022). Culturing Mouse Fetal Neural Precursor Cells in a Free-Floating Serum-Free Condition. In: Deleyrolle, L.P. (eds) Neural Progenitor Cells. Methods in Molecular Biology, vol 2389. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1783-0_1
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DOI: https://doi.org/10.1007/978-1-0716-1783-0_1
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