Characteristics of the Dissociated Spiral Ganglion Cells from the Neonatal Rats by Immunohistochemical Staining

Chung, Won Ho; Rho, Hyoung Il; Kim, Myung Soon; Chang, Byung Chan; Kim, Chang Hoon; Hong, Sung Hwa

Abstract

BACKGROUND AND OBJECTIVES: Neuronal dissociation culture is an important tool for the study of neuronal cell survival and apoptosis. To establish the culture methods, the cultured cells should be proved to have the same characteristics as the spiral ganglion neurons in vivo. This study was aimed to establish the culture systems for the spinal neuronal cells and to characterize the cultured cells using different neuronal marker.
Materials and Methods
Dissociated spiral ganglion cells were harvested from Sprague Dawley rats in postnatal 5 or 6 days and cultured for 48 hours. To prepare in vivo section, the harvested cochlea was embedded in paraffin and sectioned in 4 nm thickness. Both cultured cells and paraffin sections were labeled with several monoclonal antibodies (NSE, NF-200, NF-160, S-100) and compared by the immunohistochemical and immunoflourescent staining methods.
RESULTS
Each of the antibodies was used to stain both cultured cells and paraffin sections. NSE was used to stain the nuclei of neuronal cells. Either NS or NR was used to stain both neuronal perikarya and neurite. The Schwann cells were stained by S-100. There was no significant difference in the immunostaining pattern between cochlea tissue and dissociated cells of spiral ganglion.
CONCLUSION
A dissociated culture system for the spiral ganglions was established. In the culture system, the dissociated spiral ganglion neurons preserve the neuronal characteristics. This dissociation culture system can be used for the study of neuronal cell biology.