Abstract: Objective To determine optimal in vitro culture systems for guinea pig's geniculate ganglion cells(GGCs) and spiral ganglion cells(SGCs) by characterizing the in vitro growth of GGCs and SGCs under different culture systems. Methods Full path of facial nerve coursing was anatomized through the temporal bone and spiral ganglion of adult guinea pigs was dissected. The facial nerve and spiral ganglion tissue were digested with trypsin or collagenase typeⅠ. After enzymatic treatment, the remaining tissue was cultured with a serum-free medium supplemented with a substitute of serum or with a medium supplemented with low concentration of serum and mitotic inhibitor. The survival and appearance of GGCs and SGCs were checked under microscope. Neurons were identified by immunofluorescence method. Results Under either system we tested, GGCs and SGCs of adult guinea pig could be maintained alive successfully. In-vitro ganglion cells were able to grow into typical shape of neurons. Degeneration of ganglion cells was more significant in the medium containing serum and mitotic inhibitor. The appearance of non-neuron cells was more profound in medium supplemented with serum either. The GGCs and SGCs are identified with mouse antineutron-specific nuclear protein antibody by immunohistochemical method. Conclusion GGCs and SGCs of adult guinea pig could survive in appropriate in-vitro culture condition. Both the medium with low concentration of serum and mitotic inhibitor and the serum-free medium supplemented with a substitute of serum are able to maintain the growth of GGCs and SGCs in vitro.

Key words: geniculate ganglion, spiral ganglion, cell culture, guinea pig