Conditional immortalization of neuronal cells from postmitotic cultures and adult CNS

Eva M. Eves, John Kwon, Martha Downen, Marcy S. Tucker, Bruce H. Wainer, Marsha R. Rosner

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Abstract

To ddetermine whether postmitotic neurons can be immortalized by oncogenic transduction, we used two approaches involving conditional expression of a temperature-sensitive SV40 large T antigen (Tts). Initially, Tts was introduced into E17 rat embryonal hippocampal cells that were then cultured at the non-permissive temperature to enrich for postmitotic pyramidal neurons, and subsequently cloned at the permissive temperature. One clonal line (HMR10-3) expressed neuron-specific proteins upon differentiation, was capable of generating action potentials, and formed synapses with primary rat neurons in co-culture. Replating of these postmitotic cells at the permissive temperature resulted in reversible loss of neurofilament expression. Conditionally immortalized cell lines were also generated from the brain of an adult mouse carrying an inducible Tts transgene. These lines proliferated in a T antigen-dependent manner and expressed neuron-specific proteins upon differentiation at the non-permissive temperature. These results suggest that postmitotic neurons can be induced to enter the cell cycle without losing their commitment to a neuronal lineage.

Original languageEnglish (US)
Pages (from-to)396-404
Number of pages9
JournalBrain Research
Volume656
Issue number2
DOIs
StatePublished - Sep 12 1994

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ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

Cite this

Eves, E. M., Kwon, J., Downen, M., Tucker, M. S., Wainer, B. H., & Rosner, M. R. (1994). Conditional immortalization of neuronal cells from postmitotic cultures and adult CNS. Brain Research, 656(2), 396-404. https://doi.org/10.1016/0006-8993(94)91484-2