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

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Book cover In Vitro Neurotoxicology

Part of the book series: Methods in Molecular Biology ((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.

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References

  1. Banker, G.,(1998) Culturing Nerve Cells, ed., Massachusetts Institute of Technology, Boston, MA, pp. 9–36.

    Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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. Banker, G.,(1998) Culturing Nerve Cells, ed., Massachusetts Institute of Technology, Boston, MA, pp. 177–205.

    Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  Google Scholar 

  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.

    Article  Google Scholar 

  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. 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.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

Research by the authors is supported by grants from NIH (NIAAA, NIEHS), EPA, DoD, and NSF.

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Correspondence to Lucio G. Costa .

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© 2011 Springer Science+Business Media, LLC

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Giordano, G., Costa, L.G. (2011). Primary Neurons in Culture and Neuronal Cell Lines for In Vitro Neurotoxicological Studies. In: Costa, L., Giordano, G., Guizzetti, M. (eds) In Vitro Neurotoxicology. Methods in Molecular Biology, vol 758. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-170-3_2

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  • DOI: https://doi.org/10.1007/978-1-61779-170-3_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-169-7

  • Online ISBN: 978-1-61779-170-3

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