title: Establishment and characterization of a new cell line (SSP-9) derived from Atlantic salmon Salmo salar that expresses type I ifn.
abstract: In the present work, the establishment and biological characterization of a new cell line, SSP-9, derived from the pronephros of the Atlantic salmon Salmo salar, are reported. These cells grew well in Leibovitz's (L15) medium supplemented with 10% foetal calf serum at temperatures from 15 to 25° C, and they have been sub-cultured over 100 passages to produce a continuous cell line with an epithelial-like morphology. The SSP-9 cells attached and spread efficiently at different plating densities, retaining 80% of cell viability after storage in liquid nitrogen. When karyotyped, the cells had 40-52 chromosomes, with a modal number of 48. Viral susceptibility tests showed that SSP-9 cells were susceptible to infectious pancreatic necrosis virus and infectious haematopoietic necrosis virus, producing infectious virus and regular cytopathic effects. Moreover, these cells could be stimulated by poly I:C, showing significant up-regulation in the expression of the genes that regulate immune responses, such as ifn and mx-1. SSP-9 cells constitutively express genes characteristic of macrophages, such as major histocompatibility complex (mhc-II) and interleukin 12b (il-12b), and flow cytometry assays confirmed that SSP-9 cells can be permanently transfected with plasmids expressing a reporter gene. Accordingly, this new cell line is apparently suitable for transgenic manipulation, and to study host cell-virus interactions and immune processes.
authors.affiliation: Centro de Investigaciones Biológicas, CSIC, C/Ramiro de Maeztu 9, 28040, Madrid, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Rodriguez Saint-Jean
authors.forename: C
authors.initials: C
authors.lastname: González
authors.forename: M
authors.initials: M
authors.lastname: Monrás
authors.forename: A
authors.initials: A
authors.lastname: Romero
authors.forename: N
authors.initials: N
authors.lastname: Ballesteros
authors.forename: R
authors.initials: R
authors.lastname: Enríquez
authors.forename: S
authors.initials: S
authors.lastname: Perez-Prieto
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D015925:Cryopreservation; D007370:Interferon Type I / Q000235:genetics*; D059785:Karyotype; D060910:Pronephros / Q000166:cytology*; D020317:Salmo salar
chemical_list: D007370:Interferon Type I
keywords: fish cell line; immune genes; infectious haematopoietic necrosis virus; infectious pancreatic necrosis virus; interferon