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Establishment and characterization of an ovarian cell line from half-smooth tongue sole Cynoglossus semilaevis

文献类型: 外文期刊

作者: Sun, A. 1 ; Wang, T. Z. 1 ; Wang, N. 1 ; Liu, X. F. 1 ; Sha, Z. X. 1 ; Chen, S. L. 1 ;

作者机构: 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Marine Fisheries, Minist Agr, Qingdao 266071, Peoples R China

关键词: flatfish;gene expression;GFP transfection;karyotype analysis;ovarian granulosa cells

期刊名称:JOURNAL OF FISH BIOLOGY ( 影响因子:2.051; 五年影响因子:2.25 )

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收录情况: SCI

摘要: A new ovarian cell line, CSO, was established from half-smooth tongue sole Cynoglossus semilaevis. Primary culture of CSO cells was initiated from digestion of ovarian tissues pieces by trypsin solution and cultured at 24 degrees C in Dulbecco's modified Eagle's medium-F12 medium (DMEM-F12, 1:1) (pH 70), supplemented with 20% foetal bovine serum, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I) and human chorionic gonadotropin (HCG). The cultured CSO cells, fibroblastic in morphology, proliferated to 100% confluency 3 days later and had been subcultured to passage 80. Chromosome analyses indicated that the CSO cells exhibited chromosomal aneuploidy with a modal chromosome number of 42 that displayed the normal diploid karyotype of C. semilaevis [2n=42t, fundamental number (N-F) = 42]. Reverse transcription polymerase chain reaction revealed that CSO cells could express ovarian somatic cell functional genes p450armo, foxl2 and sox9a but not ovary germ cell marker gene vasa and male-specific gene dmrt1. Transfection experiment demonstrated that CSO cells transfected with pEGFP-N3 plasmid could express green fluorescence protein (GFP) with higher transfection efficiency. The CSO cell line might serve as a valuable tool for studies on the mechanism of sex determination and oogenesis of ovary in flatfish.

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