Establishment and Application of Blastocyst Cell Line of Olive Flounder Paralichthys olivaceus
NIE Miaomiao1, WU Zhihao2, WANG Lijuan2, YOU Feng2,3
1. State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining 810016, China; 2. State Key Laboratory of Breeding Biotechnology and Sustainable Aquaculture, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China; 3. Laboratory for Marine Biology and Biotechnology, Qingdao Marine Science and Technology Center, Qingdao 266071, China
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.
聂苗苗, 吴志昊, 王丽娟, 尤锋. 1种牙鲆囊胚期细胞系的构建及初步应用研究[J]. 水产科学, 2025, 44(4): 651-658.
NIE Miaomiao, WU Zhihao, WANG Lijuan, YOU Feng. Establishment and Application of Blastocyst Cell Line of Olive Flounder Paralichthys olivaceus. Fisheries Science, 2025, 44(4): 651-658.
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2025, Vol. 44 
