Abstract
A novel continuous ovary cell line from barfin flounder (Verasper moseri) (BFO cell line) was established with its primitive application in transgenic expression demonstrated in this study. Primarily cultured cells grew well at 22°C in Dulbecco’s modified Eagle medium/F12 medium (DMEM/F12, 1:1; pH 7.2) supplemented with 20% fetal bovine serum (FBS), carboxymethyl chitooligosaccharide, basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I). The primary BFO cells in fibroblastic morphology proliferated into a confluent monolayer about 2 weeks later, and were able to be subcultured. Impacts of medium and temperature on the growth of the cells were examined. The optimum growth was found in DMEM/F12 with 20% FBS and at 22°C. The BFO cells can be continuously subcultured to Passage 120 steadily with a population doubling time of 32.7 h at Passage 60. Chromosome analysis revealed that 72% of BFO cells at Passage 60 maintained the normal diploid chromosome number (46) with a normal karyotype of 2st+44t. The results of gene transformation indicated that green fluorescence protein (GFP) positively expressed in these cells after being transformed with pcDNA3.1-GFP. Therefore, a continuous and transformable BFO cell line was successfully established, which may serve as a useful tool for cytotechnological manipulation and transgenic modification of this fish.
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Alok, D., Hassin, S., Sampath, K. R., Trant, J. M., Yu, K., and Zohar, Y., 2000. Characterization of a pituitary GnRH-receptor from a perciform fish, Morone saxatilis: Functional expression in a fish cell line. Molecular and Cellular Endocrinology, 168: 65–75.
Aluru, N., and Vijayan, M. M., 2009. Stress transcriptomics in fish: A role for genomic cortisol signaling. General and Comparative Endocrinology, 164: 142–150.
Avella, M., Berhaut, J., and Payan, P., 1994. Primary culture of gill epithelial cells from the sea bass Dicentrarchus Labrax. In Vitro Cell and Developmental Biology–Animal, 30: 41–49.
Bols, N. C., Brubacher, J. L., Ganassin, R. C., and Lee, L. E. J., 2001. Ecotoxicology and innate immunity fish. Developmental and Comparative Immunology, 25: 853–873.
Boonstra, J., Rijken, P., Humbel, B., Cremers, F., Verkleij, A., and Bergen, E. H. P., 1995. The epidermal growth factor. Cell Biology International, 19: 413–430.
Castro, R., Martin, S. A., Zou, J., and Secombes, C. J., 2010. Establishment of an IFN-gamma specific reporter cell line in fish. Fish and Shellfish Immunology, 28: 312–319.
Chen, S. L., Ren, G. C., Sha, Z. X., and Shi, C. Y., 2004. Establishment of a continuous embryonic cell line from Japanese flounder Paralichthys olivaceus for virus isolation. Diseases of Aquatic Organisms, 60: 241–246.
Chen, S. L., Sha, Z. X., Ye, H. Q., Liu, Y., Tian, Y. S., Hong, Y., and Tang, Q. S., 2007. Pluripotency and chimera competence of an embryonic stem cell line from the sea perch (Lateolabrax japonicus). Marine Biotechnology (New York), 9: 82–91.
Dong, C., Weng, S., Li, W., Li, X., Yi, Y., Liang, Q., and He, J., 2011. Characterization of a new cell line from caudal fin of koi, Cyprinus carpio koi, and first isolation of cyprinid herpesvirus 3 in China. Virus Research, 161: 140–149.
Fan, T. J., Guo, X. Y., Jiang, G. J., Xu, X. H., Sun, A., and Xu, B., 2010a. Establishment of a continuous gill cell line from spotted halibut, Verasper variegatu. Periodical of Ocean University of China, 40: 69–74.
Fan, T. J., Jin, L. Y., and Wang, X. F., 2003. Initiation of cartilage cell culture from skate (Raja porasa Gunther). Marine Biotechnology (New York), 5: 64–69.
Fan, T. J., Xu, X. H., Jiang, G. J., Guo, X. Y., Sun, A., and Xu, B., 2010b. Establishment and identification of a contineous fin cell line from barfin flounder Verasper moser. Journal of Shandong University (Natural Science), 45: 22–27.
Fan, T., Zhao, J., Fu, Y., Cong, R., Guo, R., Liu, W., Han, B., Yu, Q., and Wang, J., 2007. Establishment of a novel corneal endothelial cell line from domestic rabbit, Oryctolagus curiculus. Science in China Series C–Life Sciences, 50: 161–169.
Goswami, M., Sharma, B, S,, Tripathi, A. K., Yadav, K., Bahuguna, S, N., Nagpure, N. S., Lakra, W. S., and Jena, J. K., 2012. Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland). Gene, 500: 140–147.
Holopainen, R., Tapiovaara, H., and Honkanen, J., 2012. Expression analysis of immune response genes in fish epithelial cells following ranavirus infection. Fish and Shellfish Immunology, 32: 1095–1105.
Huang, F., Cao, S., Cui, X., Xiong, C., Wang, M., Lu, Y., Wang, W., Ye, J., and Liu, X., 2011. Efficient gene delivery into fish cells by an improved recombinant baculovirus. Journal of Virological Methods, 173: 294–199.
Iida, J., Meijne, A. M., Oegema, T. J., Yednock, T. A., Kovach, N. L., Furcht, L. T., and McCarthy, J. B., 1998. A role of chondroitin sulfate glycosaminoglycan binding site in a4ß1 integrin-mediated melanoma cell adhesion. The Journal of Biological Chemistry, 273: 5955–5962.
Jin, L. M., Wei, C. Z., Tian, W. J., Zhao, X. J., Wang, Y. L., and Fan, S. D., 2008. Chitooligosaccharide and its derivatives promote the proliferation of osteoblasts in vitro. Journal of Clinical Rehabilitative Tissue Engineering Research, 12: 3637–3640.
Jensen, I., Steiro, K., Sommer, A. I., Mennen, S., Johansen, A., Sandaker, E. K., and Seppola, M., 2013. Establishing a cell line from Atlantic cod as a novel tool for in vitro studies. Fish and Shellfish Immunology, 34: 199–208.
Kang, M., Oh, M., Kim, Y., Kawai, K., and Jung, S., 2003. Establishment and characterization of two new cell lines derived from flounder, Paralichthys olivaceus (Temminck & Schlegel). Journal of Fish Diseases, 26: 657–665.
Kasai, H., and Yoshimizu, M., 2001. Establishment of two Japanese flounder Paralichthys olivaceus embryo cell lines. Hokkaidou Daigaku Suisan Kagaku Kenkyuu Ihou, 52: 2.
Kienzler, A., Tronchère, X., Devaux, A., and Bony, S., 2012. Assessment of RTG-W1, RTL-W1, and PLHC-1 fish cell lines for genotoxicity testing of environmental pollutants by means of a Fpg-modified comet assay. Toxicology in Vitro: An International Journal Published in Association with BIBRA, 26: 500–510.
Kim, M. S., Jee, B. Y., Cho, M. Y., Kim, J. W., Jeong, H. D., and Kim, K. H., 2012. Fugu double U6 promoter-driven long double-stranded RNA inhibits proliferation of viral hemorrhagic septicemia virus (VHSV) in fish cell lines. Archives of Virology, 157: 1029–1038.
Kojima, H., Takeuchi, S., Tsutsumi, T., Yamaguchi, K., Anezaki, K., Kubo, K., Iida, M., Takahashi, T., Kobayashi, S., Jin, K., and Nagai, T., 2011. Determination of dioxin concentrations in fish and seafood samples using a highly sensitive reporter cell line, DR-EcoScreen cells. Chemosphere, 83: 753–759.
Ku, C., Teng, Y., Wang, C., and Lu, C., 2009. Establishment and characterization of three cell lines derived from the rockfish grouper Epinephelus quoyanus: Use for transgenic studies and cytotoxicity testing. Aquaculture, 294: 147–151.
Laing, K. J., and Hansen, J. D., 2011. Fish T cells: Recent advances through genomics. Developmental and Comparative Immunology, 35: 1282–1295.
Lakra, W. S., Swaminathan, T. R., and Joy, K. P., 2011. Development, characterization, conservation and storage of fish cell lines: A review. Fish Physiology and Biochemistry, 37: 1–20.
Li, J., Sun, B., Han, H., Ouyang, J., Yao, H., Yang, Y., Liu, K., and Li, Y., 2012. Application of baculovirus as a delivery vehicle for study of transcription and translation mechanism of parvovirus in non-permissive mammalian cells. Journal of Virological Methods, 183: 1–7.
Li, W., and Li, H., 2006. Biology and culture of barfin flounder Verasper moseri in Japan. Fisheries Science, 25: 535–536.
Maclean, N., 2011. Cellular, molecular, genomics, and biomedical approaches, transgenesis and chromosome manipulation in fish. Encyclopedia of Fish Physiology, Elsevier, 1998–2008.
Meguro, Y., Nakai, T., Muroga, K., and Sorimachi, M., 1991. A cell line derived from the fin of Japanese flounder, Paralichthys olivaceus. Fish Pathology, 26: 69–75.
Miyata, T., Ishida, T., and Abe, S., 2005. A marketing study of new aquaculture, barfin flounder. Bulletin of Iwate Prefectural Fisheries Technology Center, 5: 21–33.
Parameswaran, V., Ishaq, A. V., Shukla, R., Bhonde, R. R., and Sahul, H. A., 2007. Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation. Marine Biotechnology (New York), 9: 281–291.
Pombinho, A. R., Laiz, V., Molha, D. M., Marques, S. M. P., and Cancela, M. L., 2004. Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. Cell and Tissue Research, 315: 393–406.
Qin, Q. W., Wu, T. H., Jia, T. L., Hegde, A., and Zhang, R. Q., 2006. Development and characterization of a new tropical marine fish cell line from grouper, Epinephelus coioides susceptible to iridovirus and nodavirus. Journal of Virologocal Methods, 131: 58–64.
Servili, A., Bufalino, M. R., Nishikawa, R., Melo, I. S., Muñoz-Cueto, J. A., and Lee, L. E. J., 2009. Establishment of long term cultures of neural stem cells from adult sea bass, Dicentrarchus labrax. Comparative Biochemistry and Physiology. Part A, Molecular and Integrative Physiology, 152: 245–254.
Sun, A., Fan, T., Yang, X., Xu, B., Xu, X., and Guo, X., 2011. Establishment of a spleen cell line from large yellow croaker Pseudosciaena crocea and its primitive application in foreign gene transfection. Journal of Ocean University of China, 10: 397–401.
Tafalla, C., Estepa, A., and Coll, J. M., 2006. Fish transposons and their potential use in aquaculture. Journal of Biotechnology, 123: 397–412.
Taju, G., Abdul, M. S., Nambi, K. S., and Sahul, H. A., 2013. Development and characterization of cell line from the gill tissue of Catla catla (Hamilton, 1822) for toxicological studies. Chemosphere, 90: 2172–2180.
Thomas, P., 2012. Rapid steroid hormone actions initiated at the cell surface and the receptors that mediate them with an emphasis on recent progress in fish models. General and Comparative Endocrinology, 175: 367–383.
Tong, S., Li, H., and Miao, H., 1997. The establishment and partial characterization of a continuous fish cell line FG-9307 from the gill of flounder Paralichthys olivaceus. Aquaculture, 156: 327–333.
Wang, Y., Liu, X., Xu, Y., and Wu, Y., 2009. Study on the karyotype of Verasper moseri. Progress in Fishery Sciences, 32: 6–13.
Wei, Y. B., Fan, T. J., Jiang, G. J., Xu, X. H., and Sun, A., 2010. A novel heart-cell line from brown-marbled grouper Epinephelus fuscoguttatus and its susceptibility to iridovirus. Journal of Fish Biology, 76: 1149–1158.
Yao, E., Sun, J., Liu, S., Meng, X., Sun, T., Wang, L., and Zhang, T., 2011. Technique for artificial breeding of barfin flounder Verasper moseri. Fisheries Science, 30: 22–26.
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Xu, X., Fan, T., Jiang, G. et al. Establishment and characterization of a new marine fish cell line from ovary of barfin flounder (Verasper moseri). J. Ocean Univ. China 14, 1105–1110 (2015). https://doi.org/10.1007/s11802-015-2920-6
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DOI: https://doi.org/10.1007/s11802-015-2920-6