Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 510300, Guangzhou, dong Province, China; Ocean College, Hebei Agricultural University, Qinhuangdao, 066000, China.
Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 510300, Guangzhou, dong Province, China; Sanya Tropical Fisheries Research Institute, Sanya, 572018, China; Guangdong Provincial Engineer Technology Research Center of Marine Biological Seed Industry, Guangzhou, Guangdong Province, China.
Dev Comp Immunol. 2024 Dec;161:105243. doi: 10.1016/j.dci.2024.105243. Epub 2024 Aug 13.
The yellowfin seabream (Acanthopagrus latus) is a crucial marine resource owing to its economic significance. Acanthopagrus latus aquaculture faces numerous challenges from viral diseases, but a robust in-vitro research model to understand and address these threats is lacking. Therefore, we developed a novel A. latus cell line from head kidney cells called ALHK. This study details the development, characterisation, and viral susceptibility properties of ALHK cells. This cell line primarily comprises fibroblast-like cells and has robust proliferative capacity when cultured at 28 °C in Leibovitz's L-15 medium supplemented with 10-20% foetal bovine serum. It exhibited remarkable stability after more than 60 consecutive passages and validation through cryopreservation techniques. The specificity of the ALHK cell line's origin from A. latus was confirmed via polymerase chain reaction (PCR) amplification of the cytochrome B gene, and a chromosomal karyotype analysis revealed a diploid count of 48 (2n = 48). Furthermore, the lipofection-mediated transfection efficiency using the pEGFP-N3 plasmid was high, at nearly 40%, suggesting that ALHK cells could be used for studies involving exogenous gene manipulation. In addition, ALHK cells displayed heightened sensitivity to the large mouth bass virus (LMBV), substantiated through observations of cytopathic effects, quantitative real-time PCR, and viral titration assays. Finally, the response of ALHK cells to LMBV infection resulted in differentially expressed antiviral genes associated with innate immunity. In conclusion, the ALHK cell line is a dependable in-vitro platform for elucidating the mechanisms of viral diseases in yellowfin seabream. Moreover, this cell line could be valuable for immunology, vaccine development, and host-pathogen interaction studies.
黄鳍鲷（Acanthopagrus latus）是一种具有重要经济意义的海洋资源。黄鳍鲷养殖面临着多种病毒性疾病的挑战，但缺乏用于理解和应对这些威胁的强大体外研究模型。因此，我们从头部肾脏细胞中开发了一种新的黄鳍鲷细胞系，称为 ALHK。本研究详细介绍了 ALHK 细胞的发展、特性和病毒易感性。该细胞系主要由成纤维样细胞组成，在 28°C 的 Leibovitz 的 L-15 培养基中培养，添加 10-20%胎牛血清时具有强大的增殖能力。经过 60 多次连续传代和冷冻保存技术验证后，其表现出显著的稳定性。通过细胞色素 B 基因的聚合酶链反应（PCR）扩增，证实了 ALHK 细胞系源自 A. latus 的特异性，并且染色体核型分析显示二倍体数为 48（2n = 48）。此外，使用 pEGFP-N3 质粒进行的 lipofection 介导的转染效率很高，接近 40%，这表明 ALHK 细胞可用于涉及外源基因操作的研究。此外，ALHK 细胞对大口黑鲈病毒（LMBV）表现出高度敏感性，通过观察细胞病变效应、定量实时 PCR 和病毒滴定试验得到证实。最后，ALHK 细胞对 LMBV 感染的反应导致与先天免疫相关的抗病毒基因的差异表达。总之，ALHK 细胞系是阐明黄鳍鲷病毒性疾病机制的可靠体外平台。此外，该细胞系可用于免疫学、疫苗开发和宿主-病原体相互作用研究。
