Establishment and characterization of a novel cell line from spleen of humpback grouper (Cromileptes altivelis) and its application in pathogens susceptibility

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology Pub Date : 2025-09-01 DOI:10.1016/j.dci.2025.105458

Cao-Ying Wei , Ming-Zhu Cai , Jin-Ying Zhang , Zhen-Jie Cao , Ying Wu , Chen Zhang , Min Yang , Jing-Qun Ao , Yong-Can Zhou , Yun Sun

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引用次数: 0

Abstract

The humpback grouper (Cromileptes altivelis) is a marine fish of significant commercial value and has been identified as a potential candidate for marine aquaculture. In this research, a cell line named as CAS cells was developed from the spleen tissue of the humpback grouper. This cell line has been successfully passaged for over 100 passages, with most cells exhibiting a fibroblast-like morphology. After being cryopreserved in liquid nitrogen for three months, the survival rate of the cells was over 80 %. The sequencing of the COI and 18S rRNA genes has verified that the cell line is sourced from the humpback grouper. The CAS cells exhibited vigorous proliferation in L-15 medium supplemented with 20 % FBS at 26 °C. Chromosomal analysis indicated that the CAS cells possess a diploid chromosome number of 2n = 48. Furthermore, the expression of the GFP reporter gene in CAS cells was successfully demonstrated. Toxicity assays indicated that Vibrio harveyi exhibited greater toxicity to CAS cells compared to Photobacterium damselae. P. damselae exhibited marked host cell surface adherence and induced apoptotic morphology. Moreover, experiments on viral susceptibility showed that CAS cells were sensitive to ISKNV-HN and NNV, as evidenced by cytopathic effects and viral protein production within the cells, along with an increase in the expression of immune-related genes. Consequently, our research confirms that this cell line is appropriate for in vitro genetic modification and represents a significant resource for virus isolation and the investigation of pathogenesis.

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座头石斑鱼脾脏新细胞系的建立、鉴定及其在病原菌敏感性研究中的应用。

座头石斑鱼（Cromileptes altivelis）是一种具有重要商业价值的海洋鱼类，已被确定为海洋水产养殖的潜在候选者。本研究从座头石斑鱼脾脏组织中培养了一株CAS细胞。该细胞系已成功传代超过100代，大多数细胞呈现成纤维细胞样形态。在液氮中低温保存3个月后，细胞存活率达80%以上。COI和18S rRNA基因的测序证实了该细胞系来自座头石斑鱼。在26℃条件下，添加20%胎牛血清的L-15培养基中，CAS细胞增殖旺盛。染色体分析表明，CAS细胞具有二倍体染色体数目为2n = 48。此外，还成功地证实了GFP报告基因在CAS细胞中的表达。毒性试验表明，与豆色光杆菌相比，哈维弧菌对CAS细胞的毒性更大。豆selae表现出明显的寄主细胞表面粘附和诱导的凋亡形态。此外，病毒敏感性实验表明，CAS细胞对ISKNV-HN和NNV敏感，细胞内的细胞病变效应和病毒蛋白的产生证明了这一点，同时免疫相关基因的表达增加。因此，我们的研究证实了该细胞系适合体外遗传修饰，并代表了病毒分离和发病机制研究的重要资源。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Developmental and comparative immunology 医学-动物学

CiteScore

6.20

自引率

6.90%

发文量

206

审稿时长

49 days

期刊介绍： Developmental and Comparative Immunology (DCI) is an international journal that publishes articles describing original research in all areas of immunology, including comparative aspects of immunity and the evolution and development of the immune system. Manuscripts describing studies of immune systems in both vertebrates and invertebrates are welcome. All levels of immunological investigations are appropriate: organismal, cellular, biochemical and molecular genetics, extending to such fields as aging of the immune system, interaction between the immune and neuroendocrine system and intestinal immunity.