Establishment and characterization of a head kidney cell line from large yellow croaker Pseudosciaena crocea

• Published in: Journal of fish biology 2014-05, Vol.84 (5), p.1551-1561
• Main Authors: Wang, X. H., Wang, K. R., Nie, P., Chen, X. H., Ao, J. Q.
• Format: Article
• Language: English
• Subjects: Animals; Cell Line - cytology; Chemokines, CC - metabolism; DEAD-box RNA Helicases - metabolism; Fish Proteins - metabolism; Green Fluorescent Proteins - metabolism; Head Kidney - cytology; immune stimulation; immune-related gene; Immunization; Interferon Regulatory Factors - metabolism; Interleukin-8 - metabolism; Karyotype; Lipopolysaccharides - pharmacology; Marine; marine fish; Perciformes; Poly I-C - pharmacology; Primary Cell Culture; Pseudosciaena crocea; transfection; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 0022-1112; 1095-8649
• DOI: 10.1111/jfb.12386

A continuous cell line, LYCK, derived from the head kidney of large yellow croaker Pseudosciaena crocea was established and characterized. The LYCK cell line multiplied well in Leibovitz's‐15 (L‐15) medium supplemented with 10% foetal bovine serum (FBS) at 28° C. This cell line, with a population doubling time of 29·5 h at passage 35, has been subcultured for >70 passages. Microscopically, LYCK cells were fibroblast‐like. Chromosomal analysis revealed that the modal diploid chromosome number of LYCK cells was 48, which was identical to that of the P. crocea kidney. The cellular fluorescence could be observed in LYCK cells at 48 h after being transfected with pEGFP‐N1 plasmid DNA, indicating that LYCK cells were competent for target gene expression in vitro. The expression of mRNA transcripts of the antiviral immune‐related molecules interferon regulatory factor‐3 I(ir3), interferon regulatory factor‐7 (irf7), melanoma differentiation‐associated protein 5 (mda5) was obviously up‐regulated in LYCK cells in response to the stimulation with poly (I:C), whereas the expression of mRNA transcripts of the inflammatory cytokines tumour necrosis factor‐α2 (tnfTNF‐α2), interleukin‐8 (il8), CC chemokine (lycCC) was up‐regulated by lipopolysaccharide. These results indicated that the LYCK cell line could serve as a valuable tool for studies on immune‐related gene functions in vitro.