Gjessing Mona C, Aamelfot Maria, Batts William N, Benestad Sylvie L, Dale Ole B, Thoen Even, Weli Simon C, Winton James R
Norwegian Veterinary Institute, Oslo, Norway.
US Geological Survey Western Fisheries Research Center, Seattle, Washington, United States of America.
PLoS One. 2018 Feb 14;13(2):e0191792. doi: 10.1371/journal.pone.0191792. eCollection 2018.
Gill disease in Atlantic salmon, Salmo salar L., causes big losses in the salmon farming industry. Until now, tools to cultivate microorganisms causing gill disease and models to study the gill responses have been lacking. Here we describe the establishment and characterization of two cell lines from the gills of Atlantic salmon. Atlantic salmon gill cell ASG-10 consisted of cells staining for cytokeratin and e-cadherin and with desmosomes as seen by transmission electron microscopy suggesting the cells to be of epithelial origin. These structures were not seen in ASG-13. The cell lines have been maintained for almost 30 passages and both cell lines are fully susceptible to infection by infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus (VHSV), infectious pancreatic necrosis virus (IPNV), Atlantic salmon reovirus TS (TSRV) and Pacific salmon paramyxovirus (PSPV). While infectious salmon anemia virus (ISAV) did not cause visible CPE, immunofluorescent staining revealed a sub-fraction of cells in both the ASG-10 and ASG-13 lines may be permissive to infection. ASG-10 is able to proliferate and migrate to close scratches in the monolayer within seven days in vitro contrary to ASG-13, which does not appear to do have the same proliferative and migratory ability. These cell lines will be useful in studies of gill diseases in Atlantic salmon and may represent an important contribution for alternatives to experimental animals and studies of epithelial-mesenchymal cell biology.
大西洋鲑(Salmo salar L.)的鳃病给鲑鱼养殖业造成了巨大损失。到目前为止,一直缺乏用于培养引发鳃病的微生物的工具以及研究鳃反应的模型。在此,我们描述了从大西洋鲑鳃中建立并鉴定的两个细胞系。大西洋鲑鳃细胞ASG - 10由细胞角蛋白和E - 钙黏蛋白染色阳性的细胞组成,并且通过透射电子显微镜观察到有桥粒,这表明这些细胞起源于上皮细胞。在ASG - 13中未观察到这些结构。这两个细胞系已传代培养近30次,并且对传染性造血坏死病毒(IHNV)、病毒性出血性败血症病毒(VHSV)、传染性胰腺坏死病毒(IPNV)、大西洋鲑呼肠孤病毒TS(TSRV)和太平洋鲑副粘病毒(PSPV)的感染均完全敏感。虽然传染性鲑鱼贫血病毒(ISAV)未引起明显的细胞病变效应,但免疫荧光染色显示,ASG - 10和ASG - 13细胞系中的一部分细胞可能允许感染。与ASG - 13不同,ASG - 10在体外七天内能够增殖并迁移以闭合单层中的划痕,而ASG - 13似乎不具有相同的增殖和迁移能力。这些细胞系将有助于大西洋鲑鳃病的研究,并且可能为替代实验动物以及上皮 - 间充质细胞生物学研究做出重要贡献。