A novel fish cell line derived from the brain of Chinese perch Siniperca chuatsi: development and characterization

Abstract

In this study, a continuous cell line (named as CPB) was established from Siniperca chuatsi brain and has been subcultured >140 times. CPB cell line predominantly consisted of fibroblast-like cells that could grow better in Leibovitz's L-15 supplemented with 10% foetal bovine serum at 28° C. Polymerase chain reaction amplification of 18s recombinant (r)RNA confirmed the origin of this cell line from S. chuatsi. The CPB cell line was cryopreserved at different passage levels and revived successfully with 80-90% survival. The cell line was further characterized by chromosome number and transfection. The CPB cells were highly susceptible to infectious spleen and kidney necrosis virus (ISKNV) with a titre of 6·58-6·62 log TCID50 ml(-1) and numerous ISKNV particles were observed in the cytoplasm by transmission electron microscopy. At the same time, ISKNV infection was confirmed by reverse transcriptase polymerase chain reaction, immunodot blot and individual challenge experiments. The development and characterization of a new brain cell line from S. chuatsi were described in this study and it could be used as an in vitro tool for propagation of ISKNV and gene expression studies.

New Protocol for Cell Culture to Obtain Mitotic Chromosomes in Fishes

“…In recent years, the development of new methodologies for fish cell cultures has increased following the development of the mammalian cell cultures. In general, these methodologies use a small tissue fragment to isolate the cells and, subsequently, to culture them in an adherent substratum [8,10,12,13], while other methodologies use proteolytic enzymes to isolate the cells of the tissue [14,15,16,17]. In addition, some of these methodologies have used artificial substrate to optimize and establish difficult cultures [15,16] (Table 1).…”

Immunocytochemical characterisation of neural stem‐progenitor cells from green terror cichlid Aequidens rivulatus

“…Numbers of continuous cell lines have developed teleost brain tissues because they have numerous proliferating cells (Hinsch & Zupanc, ; Kuroyanagi et al , ; Teles et al , ). Several of the cell line have characterised NSPCs (Wen et al ., 2008 a , b , 2009, 2010; Servili et al , ), yet some of the identities remain to clarify (Lai et al , ; Chi et al , ; Ku et al , ; Cheng et al , ; Hasoon et al , ; Huang et al , ; Fu et al , ). The cell lines, however, are useful for the piscine virus investigations.…”

Glutamine and glutaminolysis are required for efficient replication of infectious spleen and kidney necrosis virus in Chinese perch brain cells

“…Chinese perch brain cells (CPB) were established in our lab [32] and were propagated and maintained at 28°C in Leibovitz's L-15 medium (GIBCO, USA) supplemented with 10% fetal bovine serum (GIBCO, USA). For the glutamine depletion studies, DMEM (GIBCO, USA) with 1 g/liter D-glucose (Sigma, USA) but lacking L-glutamine and sodium pyruvate was used.…”

“…This medium was supplemented with 2% dialyzed fetal bovine serum (Hyclone, USA), 2 mM L-glutamine (GIBCO, USA) was added and used as replete medium. Infectious spleen and kidney necrosis virus isolate (ISKNV-QY) was isolated in our lab previously [32]. ISKNV was propagated in CPB cells at 28°C and its titer was determined by TCID 50 assay.…”