Establishment and characterization of a testicular Sertoli cell line from olive flounder <i>Paralichthys olivaceus</i> -- Journal of Oceanology and Limnology,2016,34(5):1054-1063

	Cite this paper:
	PENG Limin, ZHENG Yuan, YOU Feng, WU Zhihao, ZOU Yuxia, ZHANG Peijun. Establishment and characterization of a testicular Sertoli cell line from olive flounder Paralichthys olivaceus[J]. Journal of Oceanology and Limnology, 2016, 34(5): 1054-1063

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

PENG Limin^1,2, ZHENG Yuan¹, YOU Feng¹, WU Zhihao¹, ZOU Yuxia¹, ZHANG Peijun¹

1 Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences;Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China;
2 University of Chinese Academy of Sciences, Beijing 100049, China

Abstract:

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℃ 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.

Key words: olive flounder Paralichthys olivaceus|testis Sertoli cell|chromosome|FasL|transfection

Received: 2015-03-24 Revised: 2015-05-25

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Articles by ZHENG Yuan

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