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

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• 作者：Limin Peng 彭丽敏 ; Yuan Zheng 郑媛 ; Feng You 尤锋…
• 刊名：Chinese Journal of Oceanology and Limnology
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：34
• 期：5
• 页码：1054-1063
• 全文大小：806 KB
• 刊物主题：Oceanography;
• 出版者：Springer Berlin Heidelberg
• ISSN：1993-5005
• 卷排序：34

文摘

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.Keywordsolive flounder Paralichthys olivaceustestis Sertoli cellchromosomeFasLtransfection