Skip to main content
SpringerLink
Log in

Neural Differentiation of Embryonic Stem Cells

  • Protocol
Neural Stem Cells: Methods and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 198))

  • 1242 Accesses

Abstract

Differentiation of pluripotent embryonic stem (ES) cells into specific lineages is an important source of cells for implantation and gene delivery, as well as a useful model to study patterns of differentiation and gene expression during the very early development of the mammalian embryo (1). Embryonic stem cells are derived from the blastocyst inner cell mass (2,3), and remain totipotent when grown on the surface of embryonic fibroblasts or on gelatincoated substrates in the presence of leukemia inhibitory factor (LIF). ES cells appear to have unlimited proliferative capability, and, remarkably, when returned to the inner cell mass after culture and gene manipulation, resume their development and participate fully in the formation of ALL tissue types. Recently, embryonic stem cells have been derived from human blastocysts after in vitro fertilization (IVF) (4,5). Pluripotent stem cells have also been derived from human primordial germ cells (6), with obvious clinical applications.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. O’Shea, K. S. (1999) Embryonic stem cell models of development. Anat. Rec/New Anat. 257, 32–41.

    Article  Google Scholar 

  2. Evans, M. J. and Kaufman, M. H. (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156.

    Article  PubMed  CAS  Google Scholar 

  3. Martin G. R. (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. USA 78, 7634–7638.

    Article  PubMed  CAS  Google Scholar 

  4. Thomson, J. A., Itskovitz-Eldor, J., Shapiro, S. S., Waknitz, M. A., Swiergiel, J. J., Marshall, V. S., and Jones, J. M. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.

    Article  PubMed  CAS  Google Scholar 

  5. Reubinoff, B. E., Pera, M. F., Fong, C. Y., Trounson, A., and Bongso, A. (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechnol. 18, 399–404.

    Article  PubMed  CAS  Google Scholar 

  6. Shamblott, M. J., Axelman, J., Wang, S., Bugg, E. M., Littlefield, J. W., Donovan, P. J., Blumenthal, P. D., Huggins, G. R., and Gearhart, J. D. (1998) Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc. Natl. Acad. Sci. USA 95, 13,726–13,731.

    Article  PubMed  CAS  Google Scholar 

  7. Doetschman, T. C., Eistetter, H., Katz, M., Schmidt, W., and Kemler, R. (1985) The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands, and myocardium. J. Embryol. Exp. Morphol. 87, 27–45.

    PubMed  CAS  Google Scholar 

  8. Wiles, M.V. (1995) Embryonic stem cell differentiation in vitro. Methods Enzymol. 225, 900–918.

    Article  Google Scholar 

  9. Lee, S. H., Lumelsky, N., Studer, L., Auerbach, J. M., and McKay, R. D. (2000) Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells. Nat. Biotechnol. 18, 675–679.

    Article  PubMed  CAS  Google Scholar 

  10. Bain, G., Kitchens, D., Yao, M., Huettner, J. E., and Gottlieb, D. I. (1995) Embryonic stem cells express neuronal properties in vitro. Dev. Biol. 168, 342–357.

    Article  PubMed  CAS  Google Scholar 

  11. Okabe, S., Forsberg-Nilsson, K., Spiro, A. C., Segal, M., and McKay, R. D. G. (1996) Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro. Mech. Dev. 59, 89–102.

    Article  PubMed  CAS  Google Scholar 

  12. Brüstle, O., Jones, K. N., Learish, R. D., Karram, K., Choudhary, K., Wiestler, O. D., Duncan, I. D., and McKay, R. D. (1999) Embryonic stem cell-derived glial precursors: a source of myelinating transplants. Science 285, 754–756.

    Article  PubMed  Google Scholar 

  13. McDonald, J. W., Liu, X-Z., Qu, Y., Liu, S., Mickey, S.K., Turetsky, D., GottliebD I., and Choi, D.W. (1999) Transplanted embryonic stem cells survive, differentiate and promote recovery in injured rat spinal cord. Nat. Med. 5, 1410–1412.

    Article  PubMed  CAS  Google Scholar 

  14. Klug, M. G., Soonpaa, M. H., Koh, G. Y., and Field, L. J. (1996) Genetically selected cardiomyocytes from differentiating embryonic stem cells form stable intracardiac grafts. J. Clin. Invest. 98, 216–224.

    Article  PubMed  CAS  Google Scholar 

  15. O’Shea, K. S., Gratsch, T. E., Tapscott, S. J., and McCormick, M. B. (1997) Neuronal differentiation of embryonic stem (ES) cells constituitively expressing NeuroD2 or NeuroD3. Soc. Neurosci. Abstr. 23, 1144.

    Google Scholar 

  16. Li, M., Pevny, L., Lovell-Badge, R., and Smith, A. (1998) Generation of purified neural precursors from embryonic stem cells by lineage selection. Curr. Biol. 8, 971–974.

    Article  PubMed  CAS  Google Scholar 

  17. Johe, K. K., Hazel, T. G., Muller, T., Dugich-Djordjevic, M. M., and McKay, R. D. G. (1996) Single factors direct the differentiation of stem cells from the fetal and adult central nervous system. Genes Dev. 10, 3129–3140.

    Article  PubMed  CAS  Google Scholar 

  18. O’Shea, K. S. (1991) Control of neurogenesis in embryonic stem cells. J. Cell Biol. 115, 101.

    Google Scholar 

  19. Kawasaki, H., Mizuseki, K., Nishikawa, S., Kaneko, S., Kuwana, Y., Nakanishi, S., Nishikwawa S-I., and Sasai, Y. (2000) Induction of midbrain dopaminergic neurons from ES cells by stromal cell-derived inducing activity. Neuron 28, 31–40.

    Article  PubMed  CAS  Google Scholar 

  20. Gratsch, T. E., and O’Shea, K. S. (1998) Noggin and neurogenesis in embryonic stem cells. FASEB J. 12, 974.

    Google Scholar 

  21. O’Shea, K. S., Aton, S., D’Amato, C. J., and Gratsch, T. E. (1999) Embryonic stem cell derived neuroepithelial progenitor cells. Soc. Neurosci. Abstr. 25, 528.

    Google Scholar 

  22. Kilpatrick, T. J., and Bartlet, P. F. (1993) Cloning and growth of multipotential precursors: requirements for proliferation and differentiation. Neuron 10, 255–265.

    Article  PubMed  CAS  Google Scholar 

  23. Hogan, B., Beddington, R., Costantini, F., and Lacy, E. (eds.) (1994) in Manipulating The Mouse Embryo, Second Edition. Cold Spring Harbor Press, Plainview, New York, pp. 254–262.

    Google Scholar 

  24. Robertson, E. J. (ed). (1987) Teratocarcinomas and Embryonic Stem cells: A Practical Approach. Oxford: IRL Press, pp. 71–112.

    Google Scholar 

  25. Hadjantonakis, A. K., Gertsenstein, M., Ikawa, M., Okabe, M., and Nagy, A. (1998) Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mech. Dev. 76, 79–90.

    Article  PubMed  CAS  Google Scholar 

  26. Mortensen, R. M., Conner, D. A., Chao, S., Geisterfer-Lowrance, A. A., and Seidman, J. G. (1992) Production of homozygous mutant ES cells with a single targeting construct. Mol. Cell Biol. 12, 2391–2395.

    PubMed  CAS  Google Scholar 

  27. Brewer, G. J., Torricelli, J. R., Evege, E. K., and Price, P. J. (1993) Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J. Neurosci. Res. 35, 567–576.

    Article  PubMed  CAS  Google Scholar 

  28. Li, R., Gao, W.-Q., and Mather, J. P. (1996) Multiple factors control the proliferation and differentiation of rat early embryonic (Day 9) neuroepithelial cells. Endocrine 5, 205–217.

    Article  PubMed  CAS  Google Scholar 

  29. Hammarback, J. A., Palm, S. L., Furcht, L. T., and Letourneau, P. C. (1985) Guidance of neurite outgrowth by pathways of substratum-adsorbed laminin. J. Neurosci. Res. 13, 213–220.

    Article  PubMed  CAS  Google Scholar 

  30. Lopez-Coviella, I., Berse, B., Krauss, R., Thies, R. S., and Blusztajn, J. K. (2000) Induction and maintenance of the neuronal cholinergic phenotype in the central nervous system by BMP-9. Science 289, 313–316.

    Article  PubMed  CAS  Google Scholar 

  31. Wagner, J., Akerud, P., Castro, D. S., Holm, P. C., Canals, J. M., Snyder, E. Y., Perlmann, T., and Arenas, E. (1999) Induction of a midbrain dopaminergic phenotype in Nurr1-overexpressing neural stem cells by type 1 astrocytes. Nat. Biotechnol. 17, 653–659.

    Article  PubMed  CAS  Google Scholar 

  32. McWhir, J., Schnieke, A. E., Ansell, R., Wallace, H., Colman, A., Scott, A. R., and Kind, A. J. (1996) Selective ablation of differentiated cells permits isolation of embryonic stem cell lines from murine embryos with a non-permissive genetic background. Nature Genet. 14, 223–226.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI

    K. Sue O’Shea

Authors
  1. K. Sue O’Shea
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Neurosurgery, University of South Florida College of Medicine, Tampa, FL

    Tanja Zigova PhD , Paul R. Sanberg PhD, DSc  & Juan R. Sanchez-Ramos PhD, MD ,  & 

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Sue O’Shea, K. (2002). Neural Differentiation of Embryonic Stem Cells. In: Zigova, T., Sanberg, P.R., Sanchez-Ramos, J.R. (eds) Neural Stem Cells: Methods and Protocols. Methods in Molecular Biology™, vol 198. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-186-8:03

Download citation

  • DOI: https://doi.org/10.1385/1-59259-186-8:03

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-964-3

  • Online ISBN: 978-1-59259-186-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation