caa a22 4500 606229906 CHVBK 20210128101211.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10695-015-0037-x doi (NATIONALLICENCE)springer-10.1007/s10695-015-0037-x Establishment and characterization of a gonad cell line from half-smooth tongue sole Cynoglossus semilaevis pseudomale [Elektronische Daten] [Ai Sun, Song-Lin Chen, Feng-Tao Gao, Hai-Long Li, Xiao-Feng Liu, Na Wang, Zhen-Xia Sha] A new cell line was established from half-smooth tongue sole Cynoglossus semilaevis pseudomale gonad (CSPMG). Primary culture was initiated from gonad tissues pieces, and the CSPMG cells were cultured at 24°C in Dulbecco's modified Eagle medium/F12 medium (1:1) (pH7.0), supplemented with 20% fetal bovine serum, basic fibroblast growth factor, epidermal growth factor, insulin-like growth factor-I, 2-mercaptoethanol, penicillin and streptomycin. The cultured CSPMG cells, in fibroblast shape, proliferated to 100% confluency 10days later and had been subcultured to passage 109. Chromosome analyses indicated that the CSPMG cells exhibited chromosomal aneuploidy with a modal chromosome number of 42, which displayed the normal diploid karyotype of half-smooth tongue sole (2n=42t, NF=42). Reverse transcription polymerase chain reaction revealed CSPMG cells could express gonad somatic cell functional genes Sox9a, Wt1a and weakly germ cell marker gene Vasa, but not male specific gene Dmrt1. Transfection experiment demonstrated that CSPMG cells transfected with pEGFP-N3 plasmid and small RNA could express green and red fluorescence signals with high transfection efficiency. In conclusion, a continuous CSPMG cell line has been established successfully. The cell line might serve as a valuable tool for studies on the mechanism of sex determination, sex reversal and gonad development in flatfish. Springer Science+Business Media Dordrecht, 2015 Cynoglossus semilaevis nationallicence Flat fish nationallicence Pseudomale nationallicence Sertoli cells nationallicence GFP reporter gene transfection nationallicence RNA transfection nationallicence Sun Ai Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Chen Song-Lin Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Gao Feng-Tao Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Li Hai-Long Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Liu Xiao-Feng Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Wang Na Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Sha Zhen-Xia Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut Fish Physiology and Biochemistry Springer Netherlands 41/3(2015-06-01), 673-683 0920-1742 41:3<673 2015 41 10695 https://doi.org/10.1007/s10695-015-0037-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10695-015-0037-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Ai Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Chen Song-Lin Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Gao Feng-Tao Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Li Hai-Long Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Liu Xiao-Feng Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Wang Na Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Sha Zhen-Xia Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, China aut NATIONALLICENCE 773 0- Fish Physiology and Biochemistry Springer Netherlands 41/3(2015-06-01), 673-683 0920-1742 41:3<673 2015 41 10695