Establishment of a Spleen Cell Line from Large Yellow Croaker Pseudosciaena crocea and its Primitive Application in Foreign Gene Transfection

查看全文 作　　者：SUN Ai FAN Tingjun YANG Xiuxia XU Bin XU Xiaohui GUO [1]Xueyang

机构地区：[1]College of Marine Life Sciences,Ocean University of China,Qingdao 266003,P.R.China

出　　处：《Journal of Ocean University of China》索引2011年第10卷第4期,共5页

基　　金：supported by grants from the National High Technology Research and Development Program ('863' Program) of China (Grant Nos. 2006AA10A401 and 2006AA09Z406) 摘　　要：A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco's modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies. 关键词：large yellow croaker spleen cells cell line transgenic feasibility green fluorescence protein(GFP) Pseudosciaena crocea

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Establishment of a Spleen Cell Line from Large Yellow Croaker Pseudosciaena crocea and its Primitive Application in Foreign Gene Transfection

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