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Published on: July 2, 2020
Establishment and characterization of a kidney cell line from kelp grouper Epinephelus moara
Xiao-Feng Liu1,2,3, Ya-Hong Wu1, Shi-Na Wei4
1Key Laboratory for Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, CAFS, Qingdao, 266071, China.
Abstract:
A novel cell line, Epinephelus moara kidney cell line (EMK), was established from kidneys of kelp grouper E. moara. Cells were cultured at 24 °C in Leibovitz's L-15 medium (L15) supplemented with antibiotics, basic fibroblast growth factor (bFGF), foetal bovine serum (FBS) and 2-mercaptoethanol (2-ME). EMK cells, fibroblastic in morphology, proliferated to 100% confluency in 3-4 days and were subcultured for over 50 passages. The cells could grow from 18 to 30 °C, with optimal growth at 24 °C. Chromosome analysis indicated that the modal chromosome number was 48 in the cells at passage 42. Green fluorescent signals could be observed in EMK cells when the cells were transfected with pEGFP-N3 plasmid. Moreover, a significant cytopathic effect (CPE) was observed in the cells after infection with Singapore grouper iridovirus (SGIV) or nervous necrosis virus (NNV), and viral replication was confirmed by quantitative real-time PCR (qPCR). These results suggested the potential of the EMK cell line for studies of transgene and pathogenesis of SGIV and NNV.
Insights
A new Epinephelus moara kidney cell line (EMK) was developed for studying fish viruses. This cell line supports viral replication and transgene expression, showing promise for aquaculture research.
Area of Science:
- Aquaculture
- Cell Biology
- Virology
Background:
- The kelp grouper (Epinephelus moara) is an important aquaculture species.
- Establishing continuous cell lines is crucial for studying fish diseases and developing vaccines.
Purpose of the Study:
- To establish and characterize a novel cell line from Epinephelus moara kidneys.
- To evaluate the utility of this cell line for viral infection studies and transgene expression.
Main Methods:
- Primary kidney cells were cultured and established as a cell line (EMK).
- Cell growth, optimal temperature, and chromosome number were determined.
- Transfection efficiency and susceptibility to Singapore grouper iridovirus (SGIV) and nervous necrosis virus (NNV) were assessed.
Main Results:
- The EMK cell line exhibited fibroblastic morphology and stable proliferation for over 50 passages.
- Optimal growth occurred at 24°C, with a modal chromosome number of 48.
- EMK cells supported transgene expression and showed cytopathic effects and viral replication upon infection with SGIV and NNV.
Conclusions:
- The EMK cell line is a valuable tool for research on kelp grouper iridovirus and nervous necrosis virus.
- This cell line has potential applications in aquaculture research, disease diagnostics, and antiviral development.
