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Isolation and Enrichment of Defined Neural Cell Populations from Heterogeneous Neural Stem Cell Progeny

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Neural Progenitor Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1059))

Abstract

The renewable source of neural stem cells (NSCs) with multi-lineage differentiation capability towards neurons, astrocytes, and oligodendrocytes represent an ideal supply for cell therapy of central nervous system (CNS) diseases. In spite of this, the clinical use of NSCs is hampered by heterogeneity, poor neuronal cell yield, predominant astrocytic differentiation of NSC progeny and possible uncontrolled proliferation, and tumor formation upon transplantation. The ability to generate highly enriched and defined neural cell populations from the renewable source of NSCs might overcome many of these impediments and pave the way towards their successful clinical applications.

Here, we describe a simple method for NSC differentiation and subsequent purification of neuronal progenitor cells, taking advantage of size and granularity differences between neuronal cells and other NSC progeny. This highly enriched neuronal cell population provides an invaluable source of cells for both in vitro and in vivo studies.

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References

  1. Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1707

    Article  PubMed  CAS  Google Scholar 

  2. Azari H et al (2010) Isolation and expansion of the adult mouse neural stem cells using the neurosphere assay. J Vis Exp 45:e2393

    Google Scholar 

  3. Azari H et al (2011) Establishing embryonic mouse neural stem cell culture using the neurosphere assay. J Vis Exp 47:e2457

    Google Scholar 

  4. Siebzehnrubl FA et al (2011) Isolation and characterization of adult neural stem cells. Methods Mol Biol 750:61

    Article  PubMed  Google Scholar 

  5. Reynolds BA, Tetzlaff W, Weiss S (1992) A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci 12:4565

    PubMed  CAS  Google Scholar 

  6. Suslov ON et al (2002) Neural stem cell heterogeneity demonstrated by molecular phenotyping of clonal neurospheres. Proc Natl Acad Sci U S A 99:14506

    Article  PubMed  CAS  Google Scholar 

  7. Bez A et al (2003) Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Res 993:18

    Article  PubMed  CAS  Google Scholar 

  8. Taupin P (2005) Adult neurogenesis in the mammalian central nervous system: functionality and potential clinical interest. Med Sci Monit 11:RA247

    PubMed  Google Scholar 

  9. Hofstetter CP et al (2005) Allodynia limits the usefulness of intraspinal neural stem cell grafts; directed differentiation improves outcome. Nat Neurosci 8:346

    Article  PubMed  CAS  Google Scholar 

  10. Raedt R et al (2009) Unconditioned adult-derived neurosphere cells mainly differentiate towards astrocytes upon transplantation in sclerotic rat hippocampus. Epilepsy Res 87:148

    Article  PubMed  Google Scholar 

  11. Amariglio N et al (2009) Donor-Derived Brain Tumor Following Neural Stem Cell Transplantation in an Ataxia Telangiectasia Patient. PLoS Med 6:e1000029

    Google Scholar 

  12. Rozental R et al (1995) Purification of cell-populations from human fetal brain using flow cytometric techniques. Brain Res Dev Brain Res 85:161

    Article  PubMed  CAS  Google Scholar 

  13. Schmandt T et al (2005) High-purity lineage selection of embryonic stem cell-derived neurons. Stem Cells Dev 14:55

    Article  PubMed  CAS  Google Scholar 

  14. Tsagias N et al (2011) Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes. Transfus Med 21:253

    Article  PubMed  CAS  Google Scholar 

  15. Azari H et al (2011) Purification of immature neuronal cells from neural stem cell progeny. PLoS One 6:e20941

    Article  PubMed  CAS  Google Scholar 

  16. Azari H et al (2012) The neuroblast assay: an assay for the generation and enrichment of neuronal progenitor cells from differentiating neural stem cell progeny using flow cytometry. J Vis Exp 62:e3712

    Google Scholar 

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Acknowledgments

This work was supported by the Overstreet Foundation.

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Azari, H. (2013). Isolation and Enrichment of Defined Neural Cell Populations from Heterogeneous Neural Stem Cell Progeny. In: Reynolds, B., Deleyrolle, L. (eds) Neural Progenitor Cells. Methods in Molecular Biology, vol 1059. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-574-3_9

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  • DOI: https://doi.org/10.1007/978-1-62703-574-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-573-6

  • Online ISBN: 978-1-62703-574-3

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