Generation and regulation of developing immortalized neural cell lines

Methods. 1998 Nov;16(3):345-58. doi: 10.1006/meth.1998.0689.

Abstract

The genetic and environmental signals that regulate progressive lineage elaboration in the mammalian brain are poorly understood. In addition, characterization of the developmental profiles of early central nervous system (CNS) stem/ progenitor cells and analysis of the mechanisms involved in their clonal expansion, lineage restriction, and cellular maturation have been fragmentary and elusive. These seminal neurodevelopmental issues have been examined using a series of clonally derived neural stem/progenitor cell lines established by retroviral transduction of embryonic (E16.5-E17.5) murine hippocampal and cerebellar cells using temperature-sensitive alleles (A58/U19) of the simian virus (SV) 40 large tumor (T) antigen. Under conditions permissive for T-antigen expression (33 degrees C), single neural stem cells exhibited self-renewal, clonal expansion, and both symmetric and asymmetric modes of cell division. By contrast, at the nonpermissive temperature for T-antigen expression (39 degrees C), specific sets of cytokines potentiated the progressive elaboration of neuronal, oligodendroglial, and astroglial lineage species. These observations demonstrate that a spectrum of genetic and epigenetic signals and distinct cellular processes are involved in orchestrating the evolution of individual neural lineages from regional CNS stem/progenitor species. Further, the availability of conditionally immortalized neural cell lines that can be transplanted back into the mammalian brain may represent an important experimental resource for the detailed characterization of cellular and molecular mechanisms involved in the developmental sculpting, plasticity, and regeneration of the mammalian CNS.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • 3T3 Cells
  • Alleles
  • Animals
  • Antigens, Polyomavirus Transforming / metabolism
  • Brain Tissue Transplantation
  • Cell Culture Techniques / methods*
  • Cell Cycle
  • Cell Differentiation
  • Cell Line, Transformed
  • Cerebellum / metabolism
  • Epidermal Growth Factor / metabolism
  • Growth Substances
  • Hippocampus / metabolism
  • Immunohistochemistry
  • Mice
  • Neurons / cytology*
  • Receptors, Cytokine / analysis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology
  • Temperature
  • Transfection

Substances

  • Antigens, Polyomavirus Transforming
  • Growth Substances
  • Receptors, Cytokine
  • Epidermal Growth Factor