title: 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.
authors: Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61802, USA.
authors: A Ulrike
authors: AU
authors: Uebing-Czipura
authors: Harry D
authors: HD
authors: Dawson
authors: Gail
authors: G
authors: Scherba
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D019070:Cell Lineage / Q000502:physiology*; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D006564:Herpesviridae / Q000235:genetics; D006566:Herpesviridae Infections / Q000821:virology; D006801:Humans; D007150:Immunohistochemistry; D008938:Mitosis / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D009830:Olfactory Bulb / Q000166:cytology* / Q000502:physiology; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D013234:Stem Cells / Q000502:physiology*; D013552:Swine; D019098:Telomerase / Q000235:genetics; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D016271:Proto-Oncogene Proteins c-myc; D016159:Tumor Suppressor Protein p53; D019098:Telomerase