花鲈前脂肪细胞系的建立及油酸诱导分化

ESTABLISHMENT OF A PREADIPOCYTE CELL LINE OF SPOTTED SEA BASS (LATEOLABRAX MACULATUS) AND OLEIC ACID-INDUCED DIFFERENTIATION

  • 摘要: 花鲈(Lateolabrax maculatus)前脂肪细胞系为探索鱼类脂肪细胞分化和脂肪沉积的调控作用及相关分子机制提供了一种新的体外模型。研究将花鲈腹腔脂肪组织用胶原酶消化后原代培养, 再以胰酶消化后传代, 总共经过150次传代培养后, 得到花鲈前脂肪细胞系。进而, 染色体核型分析表明该细胞系长期培养仍能保持正常表型, 进一步使用线粒体cox1cytb基因鉴定该细胞系来源于花鲈。用pEGFP-C1质粒转染细胞系, 观察到明显的绿色荧光, 表明可用于研究外源基因的表达。用不同浓度的油酸培养细胞系, 3d后细胞有明显脂滴聚集, 7d后脂滴达到最大; 油红O染色表明, 400 μmol/L油酸诱导7d后细胞即可正常分化成熟。综上所述, 实验建立了花鲈前脂肪细胞系, 该细胞系形态典型、可正常分化, 为后续研究鱼类脂肪细胞分化及脂肪沉积提供了良好的实验材料。

     

    Abstract: The proadipocyte line of spotted sea bass (Lateolabrax maculatus) provides a new in vitro model for exploring the regulation of adipocyte differentiation and adipocyte deposition in fish and the related molecular mechanisms. This experiment aimed to establish a proadipose cell line for the spotted sea bass. Peritoneal adipose tissue was digested using collagenase and pancreatic enzyme, and after multiple passages, a preadipose cell line was successfully obtained following 150 times of culture cycles. karyotype analysis showed confirmed the maintenance of a normal phenotype in the cell line during long-term culture. Additionally, the mitochondrial cox1 and cytb genes were further used to identify the cell line as originating from the spotted seabass. The established cell lines were subjected to transfection with the pEGFP-C1 plasmid, resulting in observable green fluorescence, indicating their suitability for studying the expression of foreign genes. Furthermore, when cultured with different concentrations of oleic acid, the cell lines exhibited prominent lipid droplet aggregation after 3d, reaching its maximum accumulation after 7d. Oil red O staining demonstrated that the cells could differentiate and mature normally after 7d of induction with 400 μmol/L oleic acid. In summary, this experiment successfully established a preadipose cell line for the spotted sea bass, characterized by typical morphology and normal differentiation. This cell line serves as valuable experimental material for future studies on adipose cell differentiation and adipose deposition in fish.

     

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