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Establishment of Conditions for Long-Term Maintenance of Primary Embryonic Cell Cultures from Olive Flounder Paralichthys olivaceus

발생과생식 2020년 24권 3호 p.207 ~ 214
김주원, 조자영, 김동균, 남보혜, 노은수, 김봉석, 김영옥, 공희정,
소속 상세정보
김주원 ( Kim Ju-Won ) - National Institute of Fisheries Science Biotechnology Research Division
조자영 ( Cho Ja-Young ) - National Institute of Fisheries Science Biotechnology Research Division
김동균 ( Kim Dong-Gyun ) - National Institute of Fisheries Science Biotechnology Research Division
남보혜 ( Nam Bo-Hye ) - National Institute of Fisheries Science Biotechnology Research Division
노은수 ( Nho Eun-Soo ) - National Institute of Fisheries Science Biotechnology Research Division
김봉석 ( Kim Bong-Seok ) - National Institute of Fisheries Science Biotechnology Research Division
김영옥 ( Kim Young-Ok ) - National Institute of Fisheries Science Biotechnology Research Division
공희정 ( Kong Hee-Jeong ) - National Institute of Fisheries Science Biotechnology Research Division

Abstract


Primary cell culture is a sufficient method frequently used to study the cellular properties and mechanisms of isolated cells in a controlled environment. In this study, an embryonic cell line (FGBC8) derived from the blastula stages of embryos of olive flounder Paralichthys olivaceus was developed. Furthermore, conditions for optimal long-term maintenance of this primary embryonic cell culture were investigated. Morphologically, FGBC8 cells were composed primarily of epithelial-like cells. FGBC8 cells were subcultured for >160 passages over ~830 days. The doubling time of FGBC8 cells was 73.8 h, and the modal diploid chromosome number was 48. FGBC8 cells transfected with green fluorescence protein (GFP)-expression plasmid exhibited a strong signal 48 h after transfection. Consequently, we demonstrated that fish serum is a crucial supplement for the long-term survival and maintenance of comparable morphology in these primary embryonic cells. Our results can be used as a guide for primary embryonic cell cultures for other fish species and may be useful for cell biotechnological applications.

키워드

Olive flounder; Embryo; Primary cell culture; Long-term maintenance

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