Abstract: Objective: To establish a primary culture model of trigeminal neurons in newborn SD rats and provide experimental basis for the study of trigeminal nerve cytology. Methods: Twenty-four new born SD rats were selected and the trigeminal ganglion was isolated under the microscope. Neural cell suspension was prepared by trypsin and DNase digestion and mechanical blowing. The expression of neuron specific antigen NeuN and β-tubulinⅢ was identified by immunofluorescence staining. Results: Nerve cells were adherent growth, and reached protrusions. With time, the protrusions became longer and staggered network. The cells reached the best status after cultured for 3-7 days, then gradually apoptosis. The cells could culture for 15 days without ability to pass on, and the expressions of neuronal specific antigen NeuN and β-tubulinⅢ were positive. Conclusion: The combination of trypsin and DNase digestion can isolate a large number of trigeminal neurons, which provides the experimental basis for cytology research.

Key words: Trigeminal neurons, Cell culture, Immunofluorescence