Establishment and characterization of a fibroblast-like cell line from the muscle of turbot (Scophthalmus maximus L.)

被引:31
作者
Gao, Ya [1 ,2 ]
Zhou, Huihui [1 ,2 ]
Gao, Zongyu [1 ,2 ]
Jiang, Haowen [1 ,2 ]
Wang, Xuan [1 ,2 ]
Mai, Kangsen [1 ,2 ]
He, Gen [1 ,2 ,3 ]
机构
[1] Ocean Univ China, Minist Agr, Key Lab Aquaculture Nutr, 5 Yushan Rd, Qingdao 266003, Shandong, Peoples R China
[2] Ocean Univ China, Key Lab Mariculture, Minist Educ, 5 Yushan Rd, Qingdao 266003, Shandong, Peoples R China
[3] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266237, Shandong, Peoples R China
基金
国家重点研发计划;
关键词
Turbot; Muscle; Cell line; Transfection; Amino acid sensing; SOYBEAN-MEAL; FISH-MEAL; IN-VITRO; GROWTH; METABOLISM; DIETS; SUPPLEMENTATION; REPLACEMENT; RESPONSES; CULTURE;
D O I
10.1007/s10695-019-00628-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A continuous fibroblast-like cell line, TMF (turbot muscle fibroblasts), was established from juvenile turbot Scophthalmus maximus muscle with the method of trypsin digestion. It has been subcultured more than 60 passages for over 150days. The TMF cells were cultured in L-15 medium supplemented with HEPES, fetal bovine serum (FBS), GlutaMAX, and basic fibroblast growth factor (bFGF). The optimal temperature for TMF culture was 24 degrees C. TMF cells were predominantly composed of fibroblastic-like cells, and the transcription factor 4 (TCF-4) was highly expressed in TMF cells. Chromosome analysis revealed that it had a diploid chromosome number of 2n=44. The transfection efficiency achieved 54.95 +/- 6.59%, and the cell mortality rate was about 8.70% when transfected with the nucleofection method. Meanwhile, the TMF cells showed a sensitive response to amino acid levels and activation target of rapamycin (TOR) signaling pathway. These results indicate that TMF was a potential tool to explore the signal transduction of teleost in vitro.
引用
收藏
页码:1129 / 1139
页数:11
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