Pompano鱼类已在世界范围内广泛养殖。作为Carangidae家族的代表性商业海洋物种,金色的pompano(Trachinotusblochii)在中国和全世界都很受欢迎。然而,由于快速增长和高密度的水产养殖,金帕诺已经受到各种疾病的严重威胁。细胞系是体外研究的最具成本效益的资源,由于其可及性和便利性而被广泛用于生理和病理研究。在这项研究中,我们建立了一个新的永生细胞系,GPF(金绒鳍细胞)。GPF已传代超过69代10个月。形态学,使用光学和电子显微镜评估GPF的粘附和延伸过程。每3天用含有20%胎牛血清(FBS)的L-15以1:3传代GPF细胞。GPF生长的最佳条件是28°C和20%FBS浓度。18SrRNA和线粒体16SrRNA的DNA测序证实,GPF来源于金pompano。染色体分析显示,GPF的数量模式为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)表现出针对GPF的剂量依赖性细胞毒性。聚肌苷酸-聚胞苷酸(聚I:C)处理后,视黄酸诱导基因I样受体(RLR)途径基因的转录,包括mda5,mita,tbk1、irf3和irf7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)刺激上调炎症相关因子的表达,包括myd88,irak1,nfκb,IL1β,il6和cxcl10表达。据我们所知,这是使用已建立的fin细胞系对goldenpompano的免疫应答信号通路进行的首次研究。在这项研究中,我们描述了GPF细胞系对聚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。
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.