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

• Published in: Chinese journal of oceanology and limnology Vol. 34; no. 5; pp. 1054 - 1063
• Main Author: 彭丽敏 郑媛 尤锋 吴志昊 邹玉霞 张培军
• Format: Journal Article
• Language: English
• Published: Heidelberg Science Press 01.09.2016; Springer Nature B.V
• Subjects: Animal populations; Biology; Cell culture; Cell interactions; Cells; Chromosomes; COI protein; Diploids; Diploidy; Earth and Environmental Science; Earth Sciences; epidermal growth factor; FasL protein; fetal bovine serum; Fibroblast growth factor 2; Fibroblasts; Fish; Flounder; Fluorescence; Gene expression; gene expression regulation; genes; growth and development; Growth curves; Growth factors; Immunocytochemistry; Karyotypes; karyotyping; Marine; Marine fishes; Mitochondrial DNA; Nucleotide sequence; Oceanography; Paralichthys olivaceus; PCR; plasmids; Pleuronectiformes; reverse transcriptase polymerase chain reaction; Sertoli cells; Serum; Spermatogenesis; Testes; Transfection; testis Sertoli cell; olive flounder; FasL; chromosome; transfection
• ISSN: 0254-4059; 2096-5508; 1993-5005; 2523-3521
• DOI: 10.1007/s00343-016-5091-4

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. Ceils were optimally maintained at 25℃ 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 dmrtl 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 ceils 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.