Submitted to: Journal of Neuroscience Methods
Publication Type: Peer reviewed journal
Publication Acceptance Date: 1/23/2008
Publication Date: 2/13/2008
Citation: Uebing-Czipura, U., Dawson, H.D., Scherba, G. 2008. Immortalization and characterization of lineage-restricted neuronal progenitor cells derived from the procine olfactory bulb. Journal of Neuroscience Methods. 170(2)262-276. Interpretive Summary: Biomedical research benefits from the study of natural host-pathogen systems that necessitate the use of animal models since experimentation is limited in humans. In this regard, swine fulfill the need for suitable alternatives that accurately and precisely emulate many aspects of human structure and physiology. For instance, pigs are similar to humans in terms of organ size, digestive physiology, pulmonary and coronary vasculature, social behaviors, dietary habits, and propensity to obesity. Their consequential use for medical studies has provided significant advances even to the field of neurology, such as in the understanding of the neurological dysfunctions of Alzheimer’s and Parkinson’s disorders. One limitation to their utility is the lack of well defined reagents and cell lines. Herein, we describe the development and full characterization of a neuronal cell line that will be useful in advancing the pig as a model for exploring neuronal cell-pathogen interactions in cell culture. In addition, this cell line can also act as a complement to nutritional studies conducted in similar cell lines developed from humans and rodents.
Technical 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 cultured 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 cells exhibited progenitor cell-like morphology and mitotic competency based on its sustained subpassage in culture, prevalence in the cell cycle G0/G1 phase, and overall lack of cellular senescence as compared to primary cultures. The OBGF400 neuronal phenotype was indicated by the recognition of a transfected neuronal progenitor cell-specific tubulin-alpha-1 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 probably 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, this novel neuroblast cell line may be useful for exploring neuronal cell-pathogen interactions in vitro.