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建立和鉴定来源于褐牙鲆肠道的 SSI 细胞系,用于研究其对病原菌的免疫应答。

Establishment and characterization of SSI cell line from Sebastes schlegelii intestine for investigating the immune response to Pathogenic Bacteria.

作者信息

Xue Ting, Wang Yanmin, Liu Yiping, Liu Yuping, Li Chao

机构信息

School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, 266109, China.

School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, 266109, China.

出版信息

Fish Shellfish Immunol. 2024 Nov;154:109993. doi: 10.1016/j.fsi.2024.109993. Epub 2024 Oct 29.

Abstract

As a commercially valuable fish species, Sebastes schlegelii faces threats from pathogenic bacteria like Edwardsiella piscicida during aquaculture. The global host range of E. piscicida encompasses various species, yet its pathogenic mechanism remains incompletely elucidated. Cell lines offer invaluable in vitro resources for studying the pathogen pathogenicity. Here, we established and characterized a cell line derived from the intestinal tissue of the S. schlegelii, designated as SSI. SSI has undergone continuous subculturing for over 80 passages, demonstrating robust growth in DMEM supplemented with 10%-20% FBS and 20 μM HEPES at 24°C. Karyotype analysis and 18S rRNA amplification confirm its origin. SSI exhibits high transfection efficiency for exogenous DNA, making it suitable for gene expression and intestinal function analysis. E. piscicida infects SSI cells at low densities without inducing morphological changes within 6 h of infection, suggesting the potential of SSI as an in vitro model for studying E. piscicida pathogenicity. This cell line provides a valuable tool for investigating mucosal immunity and E. piscicida pathogenic mechanisms in marine fish.

摘要

作为一种有商业价值的鱼类,拟沙丁鱼在水产养殖过程中面临着爱德华氏菌等病原菌的威胁。爱德华氏菌的全球宿主范围包括多种鱼类,但它的致病机制仍不完全清楚。细胞系为研究病原体的致病性提供了非常有价值的体外资源。在这里,我们建立并鉴定了一种源自拟沙丁鱼肠道组织的细胞系,命名为 SSI。SSI 已经连续传代培养了 80 多代,在添加了 10%-20%FBS 和 20 μM HEPES 的 DMEM 中于 24°C 下生长良好。染色体核型分析和 18S rRNA 扩增证实了它的起源。SSI 对外源 DNA 的转染效率很高,适合用于基因表达和肠道功能分析。在感染后 6 小时内,低密度的爱德华氏菌感染 SSI 细胞而不引起形态变化,表明 SSI 有潜力成为研究爱德华氏菌致病性的体外模型。该细胞系为研究海洋鱼类的黏膜免疫和爱德华氏菌致病机制提供了一个有价值的工具。

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