Abstract
In the present study, a new hepatic tissue‐origin cell line from European eel Anguilla anguilla has been developed and characterized. This cell line designated EL has been maintained in Leibovitz L‐15 supplemented with 10% fetal bovine serum over 72 months, and subcultured more than 90 times. The EL cell line consisted predominantly of fibroblast‐like cells, which could survive over 100 days in vitro, and could grow at 15–32°C. The optimum temperature for growth was 27°C. The chromosome analysis revealed a modal diploid karyotype of 2n = 38. The origin of this cell line was confirmed by the 18S recombinant (r)RNA sequencing. The susceptibility test indicated significant cytopathic effects in the EL cells with regard to the Rana grylio virus and the Herpesvirus anguillae. The viral replication was confirmed by transmission electron microscopy and polymerase chain reaction analysis. Following poly (I:C) exposure, the expression levels of the immune‐related molecules interferon regulatory factor‐7 (irf7) and transforming growth factor‐β (TGF‐β) were downregulated in EL cells, whereas the expression levels of the rf3 and the cytochrome P450 (CYP450) were upregulated. All four genes were significantly upregulated following inflammation by lipopolysaccharide (LPS). These data suggested the application of EL cell line for viral identification, as well as for immunodiagnosis and pharmacological targeting.
Highlights
In the studies of aquatic virology and toxicology, fish cell lines are considered classical models and viable tools in the fields of immunology, pharmacology, and genetics (Baksi and Frazier, 1990; Kohlpoth et al, 1999; Ni Shúilleabháina et al, 2006)
The European eel Anguilla anguilla (L. 1758) is a fish species that has been traditionally used as a main food‐ consumption product in Europe (Dekker, 2018)
In the last 50 years, the abundance of A. anguilla in its original habitat has decreased over 90% (Dekker, 2004, 2018), In 2009, A. anguilla was listed CR by International Union for Conservation of Nature (IUCN), while the precious species is still threatened by various pathogens and pollutants (Yue et al, 1998; Zhang and Gui, 2007; Jakob et al, 2009; Hendriks et al, 2010; Galinier et al, 2012; Ge et al, 2012; van Beurden et al, 2012; Fichtner et al, 2013)
Summary
In the studies of aquatic virology and toxicology, fish cell lines are considered classical models and viable tools in the fields of immunology, pharmacology, and genetics (Baksi and Frazier, 1990; Kohlpoth et al, 1999; Ni Shúilleabháina et al, 2006). In the late 1990s’ and in the first decade of the present century, A. anguilla became the most productive aquacultured Anguillidae species in China. In the last 50 years, the abundance of A. anguilla in its original habitat has decreased over 90% (Dekker, 2004, 2018), In 2009, A. anguilla was listed CR (critically endangered) by International Union for Conservation of Nature (IUCN), while the precious species is still threatened by various pathogens and pollutants (Yue et al, 1998; Zhang and Gui, 2007; Jakob et al, 2009; Hendriks et al, 2010; Galinier et al, 2012; Ge et al, 2012; van Beurden et al, 2012; Fichtner et al, 2013). The disease prevention and protection of A. anguilla requires additional viable in vitro cell line tools.
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