Establishment and characterization of an ovarian cell line from half‐smooth tongue sole Cynoglossus semilaevis. Issue 1 (31st October 2014)

• Record Type: Journal Article
• Authors: Sun, A.; Wang, T. Z.; Wang, N.; Liu, X. F.; Sha, Z. X.; Chen, S. L.
• Is Part Of: Journal of fish biology. Volume 86:Issue 1(2015)
• Page Start: 46
• Page End: 59
• Publication Date: 2014-10-31
• Subjects: Fishes -- Periodicals; Fishes -- Great Britain -- Periodicals; 597
• DOI: 10.1111/jfb.12535
• Languages: English
• ISSNs: 0022-1112

Abstract

<abstract abstract-type="main" id="jfb12535-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <p id="jfb12535-para-0001">A new ovarian cell line, CSO, was established from half‐smooth tongue sole <italic>Cynoglossus semilaevis</italic>. Primary culture of CSO cells was initiated from digestion of ovarian tissues pieces by trypsin solution and cultured at 24° C in Dulbecco's modified Eagle's medium–F12 medium (DMEM–F12, 1:1) (pH 7·0), supplemented with 20% foetal bovine serum, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin‐like growth factor‐I (IGF‐I) and human chorionic gonadotropin (HCG). The cultured CSO cells, fibroblastic in morphology, proliferated to 100% confluency 3 days later and had been subcultured to passage 80. Chromosome analyses indicated that the CSO cells exhibited chromosomal aneuploidy with a modal chromosome number of 42 that displayed the normal diploid karyotype of <italic>C. semilaevis</italic> [2<italic>n</italic> = 42 t, fundamental number (<italic>N</italic><sub>F</sub>) = 42]. Reverse transcription polymerase chain reaction revealed that CSO cells could express ovarian somatic cell functional genes <italic>p</italic>450<italic>armo</italic>, <italic>foxl2</italic> and <italic>sox9a</italic> but not ovary germ cell marker gene <italic>vasa</italic> and male‐specific gene <italic>dmrt1</italic>. Transfection experiment demonstrated that CSO cells transfected with pEGFP‐N3 plasmid could express green fluorescence protein (GFP) with higher transfection efficiency. The CSO cell line might serve as a valuable tool for studies on the mechanism of sex determination and oogenesis of ovary in flatfish.</p> </abstract>