Establishment and characterization of a testicular Sertoli cell line from olive flounder Paralichthys olivaceus

被引:0
作者
Limin Peng
Yuan Zheng
Feng You
Zhihao Wu
Yuxia Zou
Peijun Zhang
机构
[1] Qingdao National Laboratory for Marine Science and Technology,Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
[2] Qingdao National Laboratory for Marine Science and Technology,Laboratory for Marine Biology and Biotechnology
[3] University of Chinese Academy of Sciences,undefined
来源
Chinese Journal of Oceanology and Limnology | 2016年 / 34卷
关键词
olive flounder ; testis Sertoli cell; chromosome; FasL; transfection;
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学科分类号
摘要
The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.
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页码:1054 / 1063
页数:9
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