Abstract
Common bottlenose dolphin (Tursiops truncatus) is a well-known cetacean species that inhabits temperate and tropical seas worldwide. Limited supply and poor quality of samples hinder the investigation of the effects of various pathogens and environmental pollutants on this cetacean species. Cultured cells are useful for experimental studies; however, no cell lines derived from cetaceans are generally available. Therefore, in this study, we established a novel kidney cell line, TK-ST, derived from T. truncatus. Primary cells exhibited the morphological characteristics of epithelial and fibroblast cells, but their immortalization and passaging resulted in a predominantly epithelial cell morphology. TK-ST was immortalized using the large T SV40 antigen and human telomerase reverse transcriptase and exhibited long-term stable cell growth. TK-ST cells are generally cultured in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum at 37°C and 5% CO2 but can also be cultured in 5–20% fetal bovine serum and several other classical media commonly used for common animal cell culture. TK-ST cells were found to be susceptible to several viruses, including the dolphin morbillivirus (most important virus in cetaceans), and exhibited cytopathic effects, facilitating the replication of the dolphin morbillivirus. Furthermore, mRNA expression levels of cytokine genes were increased in TK-ST cells after stimulation with lipopolysaccharides and poly(I:C). Therefore, the novel TK-ST cell line derived in this study can potentially be used for further in vitro studies on cetaceans.
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Acknowledgements
We would like to thank Dr. Makoto Sugiyama (Gifu University, Gifu, Japan) for providing the dolphin morbillivirus Muc strain. We are also grateful to the cooperation of the Taiji Fisheries Cooperative Union (Wakayama, Japan) and the Fisheries Agency of Japan for providing the specimens used in this study. We would also like to thank the staff of the Shinagawa Aquarium for their assistance in collecting the blood samples used in this study.
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This research was funded by a Grant-in-Aid for Scientific Research C (18K05831) from the Japan Society for the Promotion of Science (KAKENHI).
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Kaede Tashiro, Takao Segawa, and Takuya Itou conceptualized the study. Kaede Tashiro and Takuya Itou conducted the experiments. Kaede Tashiro, Taketo Futami, Miwa Suzuki, and Takuya Itou processed samples. Kaede Tashiro, Takao Segawa, Miwa Suzuki, and Takuya Itou performed the data analysis; Kaede Tashiro drafted, revised, and edited the manuscript. All authors read the manuscript and approved the final submitted manuscript for publication.
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Tashiro, K., Segawa, T., Futami, T. et al. Establishment and characterization of a novel kidney cell line derived from the common bottlenose dolphin. In Vitro Cell.Dev.Biol.-Animal 59, 536–549 (2023). https://doi.org/10.1007/s11626-023-00786-y
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DOI: https://doi.org/10.1007/s11626-023-00786-y