Skip to main content

In Vitro Culture and Analysis of Adult Hippocampal Neural Progenitors

  • Protocol
  • First Online:
Protocols for Adult Stem Cells

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 621))

Abstract

Adult hippocampal neural progenitor cell (AHNPC) culture is a useful technique for gaining insight into adult neurogenesis, studying disease, and high throughput drug screening. The ability of AHNPCs to proliferate and differentiate into the three cell lineages of the adult brain in cell culture provides the researcher a powerful platform to study the extracellular and intracellular regulatory mechanisms in a well-controlled environment. In this chapter, we describe some of the in vitro techniques necessary to study hippocampal progenitors in the adult rat. This chapter details routine culture techniques for passaging and differentiating hippocampal progenitors. We also describe techniques for analyzing the culture state, such as proliferation and expression of cell fate markers by quantitative RT-PCR and immunofluorescence.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. 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

    Article  CAS  PubMed  Google Scholar 

  2. Palmer TD, Takahashi J, Gage FH (1997) The adult rat hippocampus contains primordial neural stem cells. Mol Cell Neurosci 8:389–404

    Article  CAS  PubMed  Google Scholar 

  3. Lai K, Kaspar BK, Gage FH, Schaffer DV (2003) Sonic hedgehog regulates adult neural progenitor proliferation in vitro and in vivo. Nat Neurosci 6:21–27

    Article  CAS  PubMed  Google Scholar 

  4. Peltier J, O’Neill A, Schaffer DV (2007) PI3K/Akt and CREB regulate adult neural hippocampal progenitor proliferation and differentiation. Dev Neurobiol 67:1348–1361

    Article  CAS  PubMed  Google Scholar 

  5. Abranches E, O’Neill A, Robertson MJ, Schaffer DV, Cabral JM (2006) Development of quantitative PCR methods to analyse neural progenitor cell culture state. Biotechnol Appl Biochem 44:1–8

    Article  CAS  PubMed  Google Scholar 

  6. Bookout AL, Cummins CL, Kramer MF, Pesola JM, Mangelsdorf DJ (2006) High-throughput real-time quantitative reverse transcription PCR. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology, Wiley, New York, Unit 15.8

    Google Scholar 

Download references

Acknowledgments

This work was funded by an NSF Graduate Research Fellowship (to JP), a California Institute for Regenerative Medicine Training Grant (T1-00007), NIH T32 GM007352 (to MJR), and NSF BES-0629202.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to David V. Schaffer .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Peltier, J., Agrawal, S., Robertson, M.J., Schaffer, D.V. (2010). In Vitro Culture and Analysis of Adult Hippocampal Neural Progenitors. In: Conboy, I., Schaffer, D., Barcellos-Hoff, M., Li, S. (eds) Protocols for Adult Stem Cells. Methods in Molecular Biology™, vol 621. Humana Press. https://doi.org/10.1007/978-1-60761-063-2_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-063-2_5

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-062-5

  • Online ISBN: 978-1-60761-063-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics