Characterization of Rat Spiral Ligament Cell Line Immortalized by Adenovirus 12-Simian Virus 40 Hybrid Virus

Abstract

Objectives: Spiral ligament fibrocytes play an important role in inner ear ion homeostasis and are classified into several subtypes according to expression of specific enzymes such as Na+,K+-ATPase, Ca++-ATPase, and carbonic anhydrase. Although our understanding of the cell and molecular biology of spiral ligament fibrocytes has increased over time, access to these cells still remains a significant hurdle hindering future studies. In this study, we aimed to establish a rat spiral ligament cell line with minimal disruption of the original characteristics. Methods: The primary spiral ligament fibrocytes were exposed to adenovirus 12-simian virus 40 hybrid virus for immortalization. Karyotypic analysis was performed after stabilization of the infected cells, and the population doubling time was compared to that of the primary cell. The cell line was characterized by immunolabeling and electron microscopy. Results: We describe the establishment and characterization of a line of type I spiral ligament fibrocytes immortalized with an adenovirus 12-simian virus 40 hybrid virus. Conclusions: This cell line can be a useful research tool for investigating the role of spiral ligament fibrocytes in homeostasis and inflammation of the inner ear.