Establishment and characterization of a new cell line derived from half-smooth tongue sole <i>Cynoglossus semilaevis</i> kidney -- Journal of Oceanology and Limnology,2020,38(2):560-570

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	LOU Yanan, SUN Bin, ZHANG Lina, LI Yong, XIAO Peng. Establishment and characterization of a new cell line derived from half-smooth tongue sole Cynoglossus semilaevis kidney[J]. Journal of Oceanology and Limnology, 2020, 38(2): 560-570

Establishment and characterization of a new cell line derived from half-smooth tongue sole Cynoglossus semilaevis kidney

LOU Yanan^1,2,3, SUN Bin⁴, ZHANG Lina^1,2, LI Yong^1,2, XIAO Peng^1,2

1 Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China;
2 Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;
3 University of Chinese Academy of Sciences, Beijing 100049, China;
4 Tianjin Weishan Road Middle School, Tianjin 300222, China

Abstract:

In this study, we established a new cell line (Cynoglossus semilaevis kidney cells, CSK) from kidney of the half-smooth tongue sole (Cynoglossus semilaevis). The cells were subcultured over 1 000 days and passaged for more than 100 times. Additionally, CSK cells were optimally maintained in Dulbecco's modified Eagle medium nutrient mixture F-12 supplemented with HEPES, antibiotics, fetal bovine serum, 2-mercaptoethanol, and basic fibroblast growth factor. The optimum growth temperature for CSK cells was 25℃, and the cells showed a fibroblast-like phenotype. Chromosome analysis revealed that CSK cells had a normal diploid karyotype with 2n=42. CSK cells were susceptible to Grouper nervous necrosis virus (NNV), and cytopathic effects were observed at 3-5 days postinfection. The NNV sensitivity of CSK cells was related to the high abundance of virions in the cytoplasm, as observed by electron microscopy. Additionally, CSK cells could be successfully transfected with a green fluorescent protein reporter plasmid, and fluorescent signals were easily observed. Finally, immunocytochemistry analysis showed that CSK cells were supporting cells. Overall, we established this new cell line, which may have potential applications in the identification of viral pathogens affecting the half-smooth tongue sole.

Key words: kidney cell line|Cynoglossus semilaevis|viral multiplication

Received: 2019-03-26 Revised: 2019-05-05

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