College of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), School of Marine Biology and Fisheries, Hainan Aquaculture Breeding Engineering Research Center, Hainan Academician Team Innovation Center, Hainan University, Haikou, 570228, China.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, The Innovation Academy of Seed Design, Chinese Academy of Sciences, Wuhan, 430072, China.
Fish Shellfish Immunol. 2024 Jun;149:109568. doi: 10.1016/j.fsi.2024.109568. Epub 2024 Apr 16.
Pompano fishes have been widely farmed worldwide. As a representative commercial marine species of the Carangidae family, the golden pompano (Trachinotus blochii) has gained significant popularity in China and worldwide. However, because of rapid growth and high-density aquaculture, the golden pompano has become seriously threatened by various diseases. Cell lines are the most cost-effective resource for in vitro studies and are widely used for physiological and pathological research owing to their accessibility and convenience. In this study, we established a novel immortal cell line, GPF (Golden pompano fin cells). GPF has been passaged over 69 generations for 10 months. The morphology, adhesion and extension processes of GPF were evaluated using light and electron microscopy. GPF cells were passaged every 3 days with L-15 containing 20 % fetal bovine serum (FBS) at 1:3. The optimum conditions for GPF growth were 28 °C and a 20 % FBS concentration. DNA sequencing of 18S rRNA and mitochondrial 16S rRNA confirmed that GPF was derived from the golden pompano. Chromosomal analysis revealed that the number pattern of GPF was 48 chromosomes. Transfection experiments demonstrated that GPF could be utilized to express foreign genes. Furthermore, heavy metals (Cd, Cu, and Fe) exhibited dose-dependent cytotoxicity against GPF. After polyinosinic-polycytidylic acid (poly I:C) treatment, transcription of the retinoic acid-inducible gene I-like receptor (RLR) pathway genes, including mda5, mita, tbk1, irf3, and irf7 increased, inducing the expression of interferon (IFN) and anti-viral proteins in GPF cells. In addition, lipopolysaccharide (LPS) stimulation up-regulated the expression of inflammation-related factors, including myd88, irak1, nfκb, il1β, il6, and cxcl10 expression. To the best of our knowledge, this is the first study on the immune response signaling pathways of the golden pompano using an established fin cell line. In this study, we describe a preliminary investigation of the GPF cell line immune response to poly I:C and LPS, and provide a more rapid and efficient experimental material for research on marine fish immunology.
彭巴诺鱼在世界范围内被广泛养殖。作为鲹科家族的一种有代表性的商业海洋物种，金鲳（Trachinotus blochii）在中国和世界范围内广受欢迎。然而，由于快速生长和高密度养殖，金鲳受到各种疾病的严重威胁。细胞系是体外研究最具成本效益的资源，由于其可及性和便利性，被广泛用于生理和病理研究。在这项研究中，我们建立了一种新的永生化细胞系，GPF（金鲳鳍细胞）。GPF 已经传代了 69 代，历时 10 个月。使用光镜和电子显微镜评估了 GPF 的形态、黏附和伸展过程。GPF 细胞每 3 天用含有 20%胎牛血清（FBS）的 L-15 传代一次，传代比为 1:3。GPF 生长的最佳条件是 28°C 和 20%FBS 浓度。18S rRNA 和线粒体 16S rRNA 的 DNA 测序证实 GPF 源自金鲳。染色体分析显示，GPF 的染色体数模式为 48 条染色体。转染实验表明，GPF 可用于表达外源基因。此外，重金属（Cd、Cu 和 Fe）对 GPF 表现出剂量依赖性的细胞毒性。在用多聚肌苷酸-多聚胞苷酸（poly I:C）处理后，视黄酸诱导基因 I 样受体（RLR）途径基因，包括 mda5、mita、tbk1、irf3 和 irf7 的转录增加，诱导 GPF 细胞中干扰素（IFN）和抗病毒蛋白的表达。此外，脂多糖（LPS）刺激上调了炎症相关因子的表达，包括 myd88、irak1、nfκb、il1β、il6 和 cxcl10 的表达。据我们所知，这是首次使用建立的鳍细胞系研究金鲳的免疫反应信号通路。在这项研究中，我们描述了对 GPF 细胞系对 poly I:C 和 LPS 免疫反应的初步研究，为海洋鱼类免疫学研究提供了更快速、高效的实验材料。
