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. 2022 Jan;45(1):121-139.
doi: 10.1111/jfd.13542. Epub 2021 Oct 5.

Development and characterization of five novel cell lines from snubnose pompano, Trachinotus blochii (Lacepede, 1801), and their application in gene expression and virological studies

Nafeez Ahmed Abdul  1 , Abdul Majeed Seepoo  1 , Taju Gani  1 , Vimal Sugumar  1 , Suryakodi Selvam  1 , Badhusha Allahbagash  1 , Abdul Nazar Abdul Kuthoos  2 , Ramesh Kumar Palsamy  3 , Paknikar Kishore M  4 , Jyutika M Rajwade  4 , Sait Sahul Hameed Azeez  1
Affiliations

Development and characterization of five novel cell lines from snubnose pompano, Trachinotus blochii (Lacepede, 1801), and their application in gene expression and virological studies

Nafeez Ahmed Abdul et al. J Fish Dis. 2022 Jan.

Abstract

Five novel permanent cell lines have been established from gill, heart, kidney, eye and fin of snubnose pompano, Trachinotus blochii. They were designated as snubnose pompano gill (SPG), snubnose pompano heart (SPH), snubnose pompano kidney (SPK), snubnose pompano eye (SPE) and snubnose pompano fin (SPF), respectively. All these cell lines were characterized and cryopreserved successfully at different passage levels. Cell lines were passaged every alternate day; SPG, SPH, SPK, SPE and SPF cell lines attained passage levels of 68, 74, 82, 79 and 106, respectively, since the initiation of their development in 2019. The cell lines grew well in Leibovitz's 15 medium containing 15% foetal bovine serum at 28°C. Immunophenotyping of the cell lines revealed the presence of fibronectin and pancytokeratin. No mycoplasma contamination was found. The transfection study revealed the gene expression efficiency of these cell lines by expressing the green fluorescent protein (GFP). The authentication on origin of cell lines from T. blochii was confirmed by amplification of species-specific mitochondrial cytochrome oxidase I gene. The results showed the susceptibility of these cell lines to fish nodavirus (FNV) and tilapia lake virus (TiLV) and resistance to cyprinid herpesvirus 2 (CyHV-2). The FNV infection in the cell lines was confirmed by RT-PCR, Western blot, ELISA and immunocytochemistry, while TiLV infection was confirmed by RT-PCR assay. These results revealed that these cell lines are suitable for virological and foreign gene expression studies.

Keywords: Trachinotus blochii; cell lines; characterization; gene expression study; virus susceptibility.

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