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

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.