Document Detail


Immortalization and characterization of lineage-restricted neuronal progenitor cells derived from the porcine olfactory bulb.
MedLine Citation:
PMID:  18358537     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
A Ulrike Uebing-Czipura; Harry D Dawson; Gail Scherba
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2008-02-13
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  170     ISSN:  0165-0270     ISO Abbreviation:  J. Neurosci. Methods     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-04-18     Completed Date:  2008-07-18     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  262-76     Citation Subset:  IM    
Affiliation:
Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61802, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Newborn
Cell Aging / physiology
Cell Line
Cell Lineage / physiology*
Cells, Cultured
DNA, Complementary / biosynthesis,  genetics
Herpesviridae / genetics
Herpesviridae Infections / virology
Humans
Immunohistochemistry
Mitosis / physiology
Neurons / physiology*
Olfactory Bulb / cytology*,  physiology
Proto-Oncogene Proteins c-myc / genetics
Stem Cells / physiology*
Swine
Telomerase / genetics
Transcription, Genetic
Tumor Suppressor Protein p53 / genetics
Grant Support
ID/Acronym/Agency:
5R21AI61380-2/AI/NIAID NIH HHS; R21 AI061380-01/AI/NIAID NIH HHS; R21 AI061380-02/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/Proto-Oncogene Proteins c-myc; 0/Tumor Suppressor Protein p53; EC 2.7.7.49/Telomerase
Comments/Corrections

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