Primary Neurons in Culture and Neuronal Cell Lines for In Vitro Neurotoxicological Studies

Part of the Methods in Molecular Biology book series (MIMB, volume 758)

Abstract

Primary neuronal cultures and neuronal cell lines derived from rodents are widely used to study basic physiological properties of neurons, and represent a useful tool to study the potential neurotoxicity of chemicals. While short-term culturing of neurons can be a very straightforward process, long-term cultures of relatively pure neuronal populations require more effort. This chapter describes methods and protocols to isolate and culture primary neurons obtained from the rodent cerebellum (cerebellar granule cells), the hippocampus, and the striatum. Furthermore, culturing of rat pheochromocytoma (PC-12) cells is also described, as these cells represent a useful and widely used cell line for in vitro neurotoxicological studies.

Key words

Primary cultures Neuronal cell lines Cerebellar granule cells Hippocampal neurons Striatal neurons PC-12 cells 

References

  1. 1.
    Banker, G.,(1998) Culturing Nerve Cells, ed., Massachusetts Institute of Technology, Boston, MA, pp. 9–36.Google Scholar
  2. 2.
    Augusti-Tocco G., Sato G. (1969) Establishment of functional clonal lines of neurons from mouse neuroblastoma. Proc. Natl. Acad. Sci. USA 64, 311–5.PubMedCrossRefGoogle Scholar
  3. 3.
    Brewer G.J., Torricelli J.R., Evege E.K., Price P.J. (1993) Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J.Neurosci. Res. 35, 567–76.PubMedCrossRefGoogle Scholar
  4. 4.
    Brewer G.J. (1995) Serum-free B27/neurobasal medium supports differentiated growth of neurons from the striatum, substantia nigra, septum, cerebral cortex, cerebellum, and dentate gyrus. J Neurosci Res. 42, 674–83.PubMedCrossRefGoogle Scholar
  5. 5.
    Llinas R.R., Walton K.D., Lang E.J. (2004). “Ch. 7 Cerebellum”. in Shepherd GM. The Synaptic Organization of the Brain. New York: Oxford University Press.Google Scholar
  6. 6.
    Banker, G.,(1998) Culturing Nerve Cells, ed., Massachusetts Institute of Technology, Boston, MA, pp. 177–205.Google Scholar
  7. 7.
    Perry S.W., Norman J.P., Litzburg A., Gelbard H.A. (2004) Antioxidants are required during the early critical period, but not later, for neuronal survival. J Neurosci Res. 78, 485–92.PubMedCrossRefGoogle Scholar
  8. 8.
    Bliss T., Lømo T. (1973). Long-lasting potentiation of synaptic transmission in the dentate area of the anaesthetized rabbit following stimulation of the perforant path. J Physiol. 232, 331–56.PubMedGoogle Scholar
  9. 9.
    Megías M., Emri Z., Freund T.F., Gulyás A.I. (2001) Total number and distribution of inhibitory and excitatory synapses on hippocampal CA1 pyramidal cells. Neuroscience 102, 527–40.PubMedCrossRefGoogle Scholar
  10. 10.
    Greene L.A., Tischler A.S. (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA. 1976 73, 2424–8.CrossRefGoogle Scholar
  11. 11.
    Huettner J.E., Baughman R.W. (1986) Primary culture of identified neurons from the visual cortex of postnatal rats. J Neurosci. 10, 3044–60.Google Scholar
  12. 12.
    Junowicz E., Spencer J.H. (1973) Studies on bovine pancreatic deoxyribonuclease A. I. General properties and activation with different bivalent metals. Biochim. Biophys. Acta. 312, 72–84.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Environmental and Occupational Health SciencesUniversity of WashingtonSeattleUSA
  2. 2.Department of Human Anatomy, Pharmacology, and Forensic ScienceUniversity of Parma Medical SchoolParmaItaly

Personalised recommendations