文章摘要
郑媛,陈松林.半滑舌鳎脾脏、卵巢组织细胞离体培养的初步研究[J].水产学报,2012,36(10):1537~1543
半滑舌鳎脾脏、卵巢组织细胞离体培养的初步研究
The preliminary researches on half-smooth tongue sole spleen and ovary cells in vitro culture
投稿时间:2012-02-15  修订日期:2012-06-04
DOI:10.3724/SP.J.1231.2012.27981
中文关键词: 半滑舌鳎  脾脏  卵巢  离体培养  传代培养
英文关键词: Cynoglossus semilaevis  spleen  ovary  in vitro culture  subculture
基金项目:国家公益性行业(农业)科研专项(200903046);国家自然科学基金重点项目(31130057);山东省泰山学者工程专项
作者单位
郑媛 中国海洋大学海洋生命学院 
陈松林 中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点实验室 
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中文摘要:
      实验采用胰酶消化法,对半滑舌鳎脾脏、卵巢组织细胞进行离体培养,结果显示,脾脏、卵巢细胞能够在添加20% FBS,50 mmol/L 2-Me,100 U/mL 青霉素,100 μg/mL 链霉素,1 mmol/L丙酮酸钠,10ng/mLbFGF 的MEM完全培养基中生长,两种细胞的形态均为成纤维样,脾脏细胞可传至第9代,卵巢细胞至今已传至第11代。脾脏、卵巢细胞染色体中期分裂相均为正常的二倍体核型(2n=42),且具有异型W染色体。20%的血清浓度更适用于卵巢细胞传代培养。利用脂质体可将含有RFP荧光报告基因的质粒成功转入脾脏细胞中。
英文摘要:
      The cell lines from different organs of half-smooth tongue sole were the ideal material for researches on fish virology,genetic screening and functional analysis,etc.In this study,the researches of the spleen and ovary cells from female individual in vitro culture were developed by trypsin digestion method.The results showed that the spleen and ovary cells were cultured in minimum essential medium(MEM)supplemented with 20% fetal bovine serum(FBS),50 mmol/L 2-Mercaptoethanol(2-Me),100 U/mL penicillin,100 μg/mL streptomycin,1 mmol/L sodium pyruvate and 10 ng/mL basic fibroblast growth factor(bFGF),and microscopically,the cells were composed of fibroblast-like cells.The spleen cells were subcultured for 9 passages.Up to now,the ovary cells had been subcultured for 11 passages.Chromosome analysis revealed that the spleen and ovary cells maintained normal diploid number(2n=42),containing the heterogametic W chromosome.In subculture,the suitable concentration of FBS for the ovary cells was 20%.The spleen cells had been successfully transformed with red fluorescent protein(RFP)reporter plasmid.
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