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


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-alpha1 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.

SUBMITTER: Uebing-Czipura AU 

PROVIDER: S-EPMC2440644 | biostudies-literature | 2008 May

REPOSITORIES: biostudies-literature

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

Uebing-Czipura A Ulrike AU   Dawson Harry D HD   Scherba Gail G  

Journal of neuroscience methods 20080213 2


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  ...[more]

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