Abstract
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 150 days. 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 °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.
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Acknowledgments
This study was supported by the National Key R&D Program of China (2018YFD0900400), National Natural Scientific Foundation of China grant (31702355 and 31772860), Aoshan Talents Cultivation Program supported by Qingdao National Laboratory for Marine Science and Technology (2017ASTCP-OS12), Fundamental Research Funds for the Central Universities (201822017) to GH, and China Agriculture Research System (CARS-47-G10) to KM.
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Gao, Y., Zhou, H., Gao, Z. et al. Establishment and characterization of a fibroblast-like cell line from the muscle of turbot (Scophthalmus maximus L.). Fish Physiol Biochem 45, 1129–1139 (2019). https://doi.org/10.1007/s10695-019-00628-3
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DOI: https://doi.org/10.1007/s10695-019-00628-3