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从绿鳍马面鲀尾鳍中开发和鉴定一种新的上皮细胞系 PSF。

Development and characterization of a new epithelial cell line PSF from caudal fin of Green chromide, Etroplus suratensis (Bloch, 1790).

机构信息

National Bureau of Fish Genetic Resources Cochin Unit, CMFRI Campus, P.O. Number 1603, Kochi, India.

出版信息

In Vitro Cell Dev Biol Anim. 2010 Sep;46(8):647-56. doi: 10.1007/s11626-010-9326-y. Epub 2010 Jun 25.

Abstract

A new cell line [pearlspot fin (PSF)] has been developed from caudal fin of Etroplus suratensis, a brackish/freshwater fish cultivated in India. The cell line was maintained in Leibovitz's L-15 supplemented with 10% fetal bovine serum (FBS). The PSF cell line consisted predominantly of epithelial-like cells. The cells were able to grow at temperatures between 25 degrees C and 32 degrees C with optimum temperature of 28 degrees C. The growth rate of PSF cells increased as the FBS proportion increased from 2% to 20% at 28 degrees C with optimum growth at the concentration of 10% FBS. One marine fish virus (fish nodavirus) was tested on this cell line and found not susceptible. After confluency, the cells were subcultured with a split ratio of 1:2. The cells showed epithelial-like morphology and reached confluency on the third d after subculture. Polymerase chain reaction amplification of mitochondrial 16S rRNA and COI indicated identity of this cell line with those reported from this fish species, confirming that the cell line was of pearlspot origin. The cells were successfully cryopreserved and revived at the tenth, 25th, and 35th passages. The bacterial extracellular products from Vibrio cholerae MTCC 3904 were found to be toxic to PSF. Karyotyping analysis indicated that the modal chromosome number was 48.

摘要

已从印度养殖的半咸水/淡水鱼类苏眉(Etroplus suratensis)的尾鳍中开发出一种新的细胞系(珍珠斑点鲷细胞系)。该细胞系在添加有 10%胎牛血清(FBS)的 Leibovitz's L-15 中维持。珍珠斑点鲷细胞系主要由上皮样细胞组成。该细胞系能够在 25°C 至 32°C 之间的温度下生长,最适温度为 28°C。在 28°C 下,随着 FBS 比例从 2%增加到 20%,PSF 细胞的生长速度增加,在 10%FBS 的浓度下生长最佳。在该细胞系上测试了一种海洋鱼类病毒(鱼诺达病毒),发现其不易感。当达到汇合时,以 1:2 的比例进行传代培养。细胞表现出上皮样形态,并在传代后第 3 天达到汇合。线粒体 16S rRNA 和 COI 的聚合酶链反应扩增表明该细胞系与该鱼类种属报告的细胞系具有同一性,证实该细胞系源自珍珠斑点鲷。该细胞系成功地进行了冷冻保存,并在第 10、25 和 35 代时复苏。霍乱弧菌 MTCC 3904 的细菌细胞外产物被发现对 PSF 有毒。核型分析表明,模式染色体数为 48。

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