Abstract:
Cobia (Rachycentron canadum), a commercially important marine fish, has been used to develop a novel gill cell line, designated CG, for the first time. The CG cell line was cultured in Leibovitz\'s-15 medium with 5% fetal bovine serum (FBS) and successfully sub-cultured more than 110 passages. It underwent verification through sequencing of the mitochondrial cytochrome C oxidase subunit I (COI) gene. Optimal growth rate was achieved when the CG cell line was cultured in a medium supplemented with 5% FBS, 1% Penicillin-Streptomycin (P/S), and 5 parts per thousand (ppt) of coral sea salt water, maintained at a temperature of 27 °C. The addition of 5 ppt of salt in the growth medium suggests that this cell line could be a viable in vitro tool for marine ecosystem toxicological studies or for culturing marine parasitic microorganisms. The CG cell line was also successfully transfected using the pTurbo-GFP plasmids, showing an 18% efficiency, with observable GFP expression. Furthermore, the cell line has been effectively cryopreserved. Gene expression analysis indicated that the CG cell line exhibits responsive regulation of immune gene expression when exposured to various stimulants, highlighting its potential as an in vitro platform for immune response studies. This makes it suitable for exploring dynamic immune signaling pathways and host-pathogen interactions, thereby offering valuable insights for therapeutic development.
摘要:
Cobia(Rachycentroncanadum),一种商业上重要的海鱼,已被用于开发一种新的ill细胞系,指定CG,第一次。将CG细胞系在含有5%胎牛血清(FBS)的Leibovitz\'s-15培养基中培养,并成功继代培养超过110代。通过对线粒体细胞色素C氧化酶亚基I(COI)基因进行测序进行了验证。当CG细胞系在补充有5%FBS的培养基中培养时,获得最佳的生长速率。1%青霉素-链霉素(P/S),和5份/千份(ppt)的珊瑚海盐水,保持在27℃的温度。在生长培养基中添加5ppt盐表明,该细胞系可能是海洋生态系统毒理学研究或培养海洋寄生微生物的可行体外工具。CG细胞系也使用pTurbo-GFP质粒成功转染,显示18%的效率,与可观察到的GFP表达。此外,该细胞系已被有效地冷冻保存。基因表达分析表明,当暴露于各种兴奋剂时,CG细胞系表现出免疫基因表达的响应性调节,强调其作为免疫反应研究的体外平台的潜力。这使得它适合探索动态免疫信号通路和宿主-病原体相互作用,从而为治疗发展提供有价值的见解。