Atlantic salmon endothelial cells from the heart were more susceptible than fibroblasts from the bulbus arteriosus to four RNA viruses but protected from two viruses by dsRNA pretreatment

Pham, Phuc H.; Tong, Winnie W.L.; Misk, Ehab; Jones, Ginny; Lumsden, John S.; Bols, Niels C.

doi:10.1016/j.fsi.2017.09.001

Cited by 9 publications

(3 citation statements)

References 92 publications

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“…The Atlantic salmon cardiovascular system has been the source for a heart cell line and bulbus arteriosus for a fibroblast cell line (Pham et al . 2017a,b). In the past decade, six macrophage cell lines were developed from fish.…”

Section: Diversity Of Fish Cell Linesmentioning

confidence: 99%

Comprehensive update on inventory of finfish cell lines developed during the last decade (2010–2020)

Thangaraj

Narendrakumar

Geetha

et al. 2021

Reviews in Aquaculture

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Cell culture has become an indispensable part of modern research and is considered the best alternatives for different biological research on whole animals as they mimic the host cellular genetic homogeneity and also reduces the variability of in vivo responses. Hence, there has been increasing development of cell lines by different research groups and companies globally. For this reason, to cover the dearth of knowledge of newly developed fish cell lines, we have made a comprehensive update on new fish cell lines established in the past decade (2010)(2011)(2012)(2013)(2014)(2015)(2016)(2017)(2018)(2019)(2020) to reflect the number of cell lines now available. This review focuses on the appraisal of the literature and of work done in finfish cell culture around the globe during the last decade and summarizes 280 newly established fish cell lines derived from 20 different tissue types of 44 fish families belonging to 21 different countries raising the total finfish cell lines to 783. The highest number of cell lines developed in the past decade was from the fish of the family Cyprinidae, which are most widely cultured in Asian countries. Also, the review discusses the need of new cell line development, different methods of cell line development and further enumerates the various applications and future prospects for these cell lines which we believe will serve as a referral document for researchers in the field.

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Section: Diversity Of Fish Cell Linesmentioning

confidence: 99%

Comprehensive update on inventory of finfish cell lines developed during the last decade (2010–2020)

Thangaraj

Narendrakumar

Geetha

et al. 2021

Reviews in Aquaculture

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“…Membranes were then blocked with 5% skim milk in PBS-T at 4°C overnight. The rest of the procedures were performed as previously described [46] and all wash steps were performed with PBS-T. The primary antibodies were either the anti-N protein monospecific rabbit antibody diluted 1:1000 or mouse monoclonal anti-beta actin diluted 1:1000 (Ther-moFisher).…”

Section: Protein Extraction Quantification and Western Blot Analysismentioning

confidence: 99%

Isolation of Ontario aquatic bird bornavirus 1 and characterization of its replication in immortalized avian cell lines

Pham

Leacy

Deng

et al. 2020

Virol J

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Background Aquatic bird bornavirus 1 (ABBV-1) has been associated with neurological diseases in wild waterfowls. In Canada, presence of ABBV-1 was demonstrated by RT-qPCR and immunohistochemistry in tissues of waterfowls with history of neurological disease and inflammation of the central and peripheral nervous tissue, although causation has not been proven by pathogenesis experiments, yet. To date, in vitro characterization of ABBV-1 is limited to isolation in primary duck embryo fibroblasts. The objectives of this study were to describe isolation of ABBV-1 in primary duck embryonic fibroblasts (DEF), and characterize replication in DEF and three immortalized avian fibroblast cell lines (duck CCL-141, quail QT-35, chicken DF-1) in order to evaluate cellular permissivity and identify suitable cell lines for routine virus propagation. Methods The virus was sequenced, and phylogenetic analysis performed on a segment of the N gene coding region. Virus spread in cell cultures, viral RNA and protein production, and titres were evaluated at different passages using immunofluorescence, RT-qPCR, western blotting, and tissue culture dose 50% (TCID50) assay, respectively. Results The isolated ABBV-1 showed 97 and 99% identity to European ABBV-1 isolate AF-168 and North American ABBV-1 isolates 062-CQ and CG-N1489, and could infect and replicate in DEF, CCL-141, QT-35 and DF-1 cultures. Viral RNA was detected in all four cultures with highest levels observed in DEF and CCL-141, moderate in QT-35, and lowest in DF-1. N protein was detected in western blots from infected DEF, CCL-141 and QT-35 at moderate to high levels, but minimally in infected DF-1. Infectious titre was highest in DEF (between approximately 105 to 106 FFU / 106 cells). Regarding immortalized cell lines, CCL-141 showed the highest titre between approximately 104 to 105 FFU / 106 cells. DF-1 produced minimal infectious titre. Conclusions This study confirms the presence of ABBV-1 among waterfowl in Canada and reported additional in vitro characterization of this virus in different avian cell lines. ABBV-1 replicated to highest titre in DEF, followed by CCL-141 and QT-35, and poorly in DF-1. Our results showed that CCL-141 can be used instead of DEF for routine ABBV-1 production, if a lower titre is an acceptable trade-off for the simplicity of using immortalized cell line over primary culture.

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“…At days 1, 3, and 5 post-inoculation, protein samples were extracted using radioimmunoprecipitation assay buffer (RIPA buffer) and stored at −20 °C until used. Detailed protein extraction, protein quantification, SDS-PAGE, and Western blot procedures were done as previously described [ 61 ]. The primary antibodies were a polyclonal rabbit antibody against rainbow trout Mx proteins (1:2000 dilution in 5% skim milk/TBS-T) and a polyclonal rabbit anti-β-actin antibody (1:1000 dilution in 5% skim milk/TBS-T; Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Screening of Fish Cell Lines for Piscine Orthoreovirus-1 (PRV-1) Amplification: Identification of the Non-Supportive PRV-1 Invitrome

et al. 2020

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Piscine reovirus (PRV) is the causative agent of heart and skeletal muscle inflammation (HSMI), which is detrimental to Atlantic Salmon (AS) aquaculture, but so far has not been cultivatable, which impedes studying the disease and developing a vaccine. Homogenates of head kidney and red blood cells (RBC) from AS in which PRV-1 had been detected were applied to fish cell lines. The cell lines were from embryos, and from brain, blood, fin, gill, gonads, gut, heart, kidney, liver, skin, and spleen, and had the shapes of endothelial, epithelial, fibroblast, and macrophage cells. Most cell lines were derived from the Neopterygii subclass of fish, but one was from subclass Chondrostei. Cultures were examined by phase contrast microscopy for appearance, and by quantitative polymerase chain reaction (qPCR) for PRV-1 RNA amplification and for the capacity to transfer any changes to new cultures. No changes in appearance and Ct values were observed consistently or transferable to new cultures. Therefore, 31 cell lines examined were unable to support PRV-1 amplification and are described as belonging to the non-supportive PRV-1 invitrome. However, these investigations and cell lines can contribute to understanding PRV-1 cellular and host tropism, and the interactions between virus-infected and bystander cells.

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Atlantic salmon endothelial cells from the heart were more susceptible than fibroblasts from the bulbus arteriosus to four RNA viruses but protected from two viruses by dsRNA pretreatment

Cited by 9 publications

References 92 publications

Comprehensive update on inventory of finfish cell lines developed during the last decade (2010–2020)

Comprehensive update on inventory of finfish cell lines developed during the last decade (2010–2020)

Isolation of Ontario aquatic bird bornavirus 1 and characterization of its replication in immortalized avian cell lines

Screening of Fish Cell Lines for Piscine Orthoreovirus-1 (PRV-1) Amplification: Identification of the Non-Supportive PRV-1 Invitrome

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