石鲽鳍细胞系的建立及三丁基锡对其毒性作用的研究

为建立石鲽鳍细胞系并研究三丁基锡对其毒性作用,根据石鲽鳍组织的特点采用特定酶消化法启动原代培养,并成功传代。试验结果显示,在24℃,培养于含有碱性成纤维样生长因子及20%胎牛血清的DMEM/F12培养液(pH值7.2)中的鳍细胞,生长分裂状态佳,为成纤维样细胞。传代培养后继续保持旺盛的生长分裂速度,可稳定传代。第60代石鲽鳍细胞的群体倍增时间为46.7 h,特征性染色体数目为48条。目前鳍细胞已传至第68代,已成功建立石鲽鳍细胞系。不同浓度三丁基锡处理后结果显示,1～80 ng/mL三丁基锡均可对鳍细胞产生明显的毒性作用,三丁基锡对鳍细胞的48 h-IC50值为39.87 ng/mL。

Establishment of Fin Cell Lines from Stone Flounder,Kareius Bicoloratus and Cytotoxic Effects of Tributyltin Chloride to the Fin Cells

To establish novel cell lines from stone flounder,Kareius bicoloratus,for the in vitro investigations of cytotoxic effects ot tributyltin chloride ,primary cultures of fin tissues were initiated by digestion methods.These ceils were successfully cultured at 24 ℃ by using 20% FBS-DMEM/ F12 medium（pH value 7.2）which was further supplemented with basic fibroblast growth factor（bFGF）.Tbe fin cells in primary culture were all in fibroblastie morphology.These cells grew at a steady rate during subsequent subculture and had a population doubling time of 46.7 h at passage 60. Karyotype analysis showed that these cells exhibited chromosomal aneuploidy with a modal chromosome number of 48.To date,the novel stone flounder fin cell lines have been established and subeultured more than passage 68.In the tributyhin chloride treated cells from 1-80 ng/mL,results showed that the 48 h-IC50 values of tributyhin chloride to stone flounder fin cells were 39.87 ng/mL.