A method for the isolation of purified murine neuroepithelial cells from the developing mouse brain

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Abstract

The adult mammalian central nervous system develops from the pseudostratified neuroepithelium of the neural tube. In order to study, in vitro, the differentiation of the neuroepithelial cells in detail and to identify factors that may influence this process, an uncontaminated, viable population of neuroepithelial cells, that still retains full developmental potential, is required. In this paper we describe a highly efficient method, involving differential trypsinization and micro-dissection, to cleanly separate the neuroepithelium from surrounding mesenchyme and ectoderm. The purity of isolated neuroepithelium has been assessed by monitoring for the presence of endothelial cells using an anti-endothelial antibody, MTS-12, and found to contain no significant level of contamination. Neuroepithelial cells prepared by this method have been demonstrated to divide and differentiate in tissue culture, to act as target cells for immortalization by proto-oncogenes and to differentiate into neurons in neural transplantation studies.

References (16)

  • AbneyE.R. et al.

    Dev. Biol.

    (1981)
  • AizenmanY. et al.

    Brain Res.

    (1987)
  • BartlettP.F. et al.

    Progr. Brain Res.

    (1990)
  • DragoJ. et al.

    Exp. Cell Res.

    (1991)
  • GensburgerC. et al.

    FEBS Lett.

    (1987)
  • GospodarowiczD. et al.

    Cell Diff.

    (1986)
  • LuskinM.B. et al.

    Neuron

    (1988)
  • RehT.A. et al.

    Dev. Biol.

    (1988)
There are more references available in the full text version of this article.

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