【摘要】 采用研磨杵压破胚胎外膜、分离出细胞，采用DF12培养基，并添加抗生素、胎牛血清和生长因子等，在25℃下从牙鲆囊胚中构建了1种多突起且边缘清晰的新细胞系，命名为PoEC,目前已经培养至40代。线粒体细胞色素氧化酶亚基Ⅰ(COⅠ)基因片段测序确定该细胞系来源于牙鲆。该细胞系具有典型的生长曲线。根据相关基因和酶活性结果，该细胞系不具有全能性。绿色荧光蛋白基因能够成功地转染该细胞系细胞，说明该细胞系可以进行牙鲆或其他鱼类基因功能研究。更重要的是，采用10、20、30、40 mg/L重金属铬处理细胞，探究重金属对细胞的影响，发现低质量浓度铬(10 mg/L)处理1 h内可以严重影响其存活率。所以该细胞系可以作为鱼类研究重金属毒理的有力工具，以体外生物学系统应用于海水鱼类生物技术和毒理学研究中。更多还原

【Abstract】 One new embryonic multipolar morphology cell line with a clearly defined birefringent edge was established by crushing the embryos with a sterile mortar to break the membrane and carefully squeezing the embryos vertically to separate the cells as much as possible, derived from blastocysts of olive flounder Paralichthys olivaceus, designated as PoEC. The cells were cultured in DF12 complete medium, supplemented with antibiotics, fetal bovine serum(FBS), and growth factors at temperature of 25 ℃, and subcultured for 40 passages. The origin of the cell lines shwoing typical growth curve pattern, was confirmed by examining the partial sequences of the cytochrome oxidase c subunit Ⅰ(COⅠ) gene of the flounder mitochondrial DNA(mtDNA). According to the gene and activity, the cell line PoEC did not have pluripotent, whose the cell line were also successfully transfected with the green fluorescent protein(GFP) reporter gene, suggesting that it was used to investigate the gene function in the flounder or other fish. The PoEC cell line was exposed to Cr concentrations of 0, 10, 20, 30, and 40 mg/L in vitro toxicity test for 16 h, and the results showed even at concentration of 10 mg/L, Cr was toxic to the cell line in one hour, PoEC being sensitive to chromium(Cr),and used as a powerful tool for research of the toxicology of heavy metals in fish. Therefore, the cell line would be useful for biotechnological and toxicological research on marine fish as an in vitro biological system.更多还原