Immortalization and characterization of lineage-restricted neuronal progenitor cells derived from the porcine olfactory bulb

A. Ulrike Uebing-Czipura, Harry D. Dawson, Gail Scherba

Research output: Contribution to journalArticlepeer-review

Abstract

Crucial aspects in the development of in vitro neuropathogenic disease model systems are the identification, characterization and continuous mitotic expansion of cultured neuronal cells. To facilitate long-term cultivation, we immortalized porcine olfactory neuronally restricted progenitor cells by genomic insertion of a cDNA encoding the catalytic subunit of the human telomerase reverse transcriptase (hTERT) yielding a stable neuroblast subclone (OBGF400). The altered cells exhibited progenitor-cell-like morphology and mitotic competency based on sustained subpassaging, prevalence in the cell cycle G0/G1 phase and an overall lack of cellular senescence as compared to primary cultures. An OBGF400 neuronal phenotype was indicated by the recognition of a transfected neuronal progenitor-cell-specific tubulin-α1 gene promoter, intracellular presence of early neuronal markers (TuJ1, neuregulin-1, doublecortin and SOX2) and enhanced expression of neuronal- and progenitor lineage-active genes (MAP2, nestin, ENO and Syn1) compared to that of porcine epithelial cells. These OBGF400 neuroblasts are likely dependent on telomerase to prevent terminal differentiation as subcultures with a predominance of neuronally differentiated members had less enzymatic activity. Based on its susceptibility to a porcine alphaherpesvirus infection, this novel neuroblast cell line may be useful for exploring neuronal cell-pathogen interactions in vitro.

Original languageEnglish (US)
Pages (from-to)262-276
Number of pages15
JournalJournal of Neuroscience Methods
Volume170
Issue number2
DOIs
StatePublished - May 30 2008

Keywords

  • Alphaherpesvirus
  • Neural transcriptome analysis
  • Olfactory bulb
  • Porcine neuroblasts
  • Porcine neuronal progenitor cells
  • hTERT

ASJC Scopus subject areas

  • Neuroscience(all)

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