Long-term culturing of porcine nodose ganglia.

ABSTRACT

BACKGROUND: Neuronal cell cultures are widely used in the field of neuroscience. Cell dissociation allows for the isolation of a desired cell type, yet the neuronal complexity that distinguishes the nervous system is often lost as a result. Thus, culturing neural tissues in ex vivo format provides a physiological context that more closely resembles the in vivo environment. NEW METHOD: We developed a simple method to culture nodose ganglia neurons from neonatal pigs long-term in ex vivo format using an in-house media formulation derived from commercially available components. RESULTS: Ganglia were cultured for six and twelve months. mRNA expression of nestin was stable across time. Vasoactive intestinal peptide and tachykinin showed statistically insignificant increases and decreases in mRNA expression, respectively. mRNA expression of glia fibrillary acidic protein decreased, whereas myelin basic protein showed no statistically significant differences, over time. Immunofluorescence studies of sectioned ganglia demonstrated neurofilament-positive cell bodies, glia fibrillary acidic protein and myelin basic protein at all time points. A significant decrease in cell nuclei density and fragmented DNA were noted. COMPARISON WITH EXISTING METHOD(S) There are currently no methods that describe short-term or long-term culturing of porcine nodose ganglia. Further, the media formulation we developed is new and not previously reported. CONCLUSIONS: The simple procedure we developed for culturing nodose ganglia will enable both short-term and long-term investigations aimed at understanding peripheral ganglia in vitro. It is also possible that the methods described herein can be applied to other models, different developmental stages, and potentially other neural tissues.