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

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

olivaceus

PENG Limin;ZHENG Yuan;YOU Feng;WU Zhihao;ZOU Yuxia;ZHANG Peijun

【期刊名称】《中国海洋湖沼学报（英文版）》 【年(卷),期】2016(034)005

【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℃ 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

\

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