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 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. Nodose ganglia neurons have been extensively studied both in dissociated form and acutely in slice format. However, methods to culture long-term ex vivo have not been established. 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 Cultures were viable for approximately 12 months. mRNA expression for nestin, a marker of neural progenitor cells, was stable across time. Vasoactive intestinal peptide and tachykinin, markers of nodose neurons, showed either no statistically significant differences or decreased across time, respectively. mRNA expression for glia fibrillary acidic protein and myelin basic protein showed no statistically significant differences over time. Comparison with Existing Method(s) There are currently no methods that describe 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 animal models, adult samples, and other neural tissues.