Development and Characterisation of an Immortal Cell Line From Largemouth Bass (Micropterus salmoides) for Viral Studies.

Autor: Mai Q; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, China., Wang Q; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Li Y; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Yin J; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Mo X; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Shi C; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Ren Y; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Liu D; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Liu X; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China., Sun D; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, China., Liu W; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, China., Jin Y; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, China., Zeng W; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, China., Wang Y; Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.
Jazyk: angličtina
Zdroj: Journal of fish diseases [J Fish Dis] 2024 Dec 24, pp. e14071. Date of Electronic Publication: 2024 Dec 24.
DOI: 10.1111/jfd.14071
Abstrakt: Largemouth bass (Micropterus salmoides), a freshwater fish species, is cultivated widely across China. The industry has been greatly affected by various viral diseases. We generated a new immortal cell line from the fin of M. salmoides (MSF). It has been successfully maintained in continuous culture for over 80 passages. Cells multiply well at 28°C using Medium 199 or Leibovitz's L-15 supplemented with 10%-20% fetal bovine serum. MSF cells comprise a mass of epithelialoid cells. Chromosome analysis showed that ploidy at passage 55 was 2n = 58. Amplification and analysis of the sequence of cytochrome oxidase I gene ascertained the origin of MSF cells from M. salmoides. The cell line is devoid of contamination with mycoplasma. Successful transfection with a GFP reporter gene indicated that the MSF cell line can be a useful tool for further gene expression studies. MSF cells exhibited susceptibility to Largemouth bass ranavirus (LMBV), Infectious spleen and kidney necrosis virus (ISKNV), and Siniperca chuatsi rhabdovirus (SCRV). Transmission electron microscopy (TEM), and immunofluorescence confirmed that the MSF cell line showed high susceptibility to all the tested viruses. The titre values of MSF after infection with LMBV, ISKNV, and SCRV reached 10 7.4 TCID 50 /mL, 10 6.15 TCID 50 /mL, and 10 6.35 TCID 50 /mL at 7dpi, respectively. In conclusion, an immortal cell line derived from the fin tissue of M. salmoides was successfully developed and certain fish viruses can effectively propagated in the newly established MSF cell line that can be helpful for future gene manipulation and infectious viral mechanistic studies in the future.
(© 2024 John Wiley & Sons Ltd.)
Databáze: MEDLINE
Externí odkaz: