Development and characterization of a new fish cell line from Honeycomb grouper, Epinephelus merra
336 / 57
Keywords:
Epinephelus merra, Fish cell line, HGF, Honeycomb grouperAbstract
A new cell line HGF was developed from fin tissue of Epinephelus merra Bloch, 1793. The cell line was maintained in Leibovitz’s L-15 supplemented with 15% FBS and cells have been subcultured 45 times. The HGF cell line consists predominantly of fibroblastic-like cells. The cells were able to grow at temperatures between 25° and 32°C with optimum temperature of 28°C. The growth rate of fin cells increased as the FBS proportion increased from 2 to 20% at 28°C with optimum growth at the concentrations of 15 or 20% FBS. After confluence, the cells were sub-cultured with a split ratio of 1:2. The cells showed fibroblastic-like morphology and reached confluence on the fourth day after subculture. Polymerase chain reaction amplification of mitochondrial 16S rRNA and COI indicated identity of these cell lines with those reported from this animal species, confirming that the cell lines were of honeycomb grouper origin. The cells were successfully cryopreserved and revived at passage numbers 10, 20 and 30. The bacterial extracellular products from Vibrio cholerae MTCC 3904 was found toxic to this cell line.
Downloads
References
Ahmed V P I, Chandra V, Sudhakaran R, Rajesh Kumar S, Sarathi M, Sarath Babu V, Bhonde R R and Sahul Hameed A S. 2009. Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton); and catla, Catla catla (Hamilton). Journal of Fish Diseases 32: 211–18.
Chang S F, Ngoh G H, Kueh L F S, Qin Q M, Che C L, Lam T J and Sin Y M. 2001. Development of a tropical marine fish cell line from Asian seabass (Lates calcarifer) for virus isolation. Aquaculture 192: 133–45.
Chi S C, Hu W W and Lo B J. 1999. Establishment and characterization of a continuous cell line (GF-1) derived from grouper; Epinephelus coioides: a cell line susceptible to grouper nervous necrosis virus (GNNV). Journal of Fish Diseases 22: 173–82.
CIBA. 2003. Training manual on sea bass (Lates calcarifer) seed production and culture. Pp 103. (Ed.) Thirunavukkarasu A R. CIBA, Chennai.
Fernandez R D, Yoshimizu M, Kimura T, Ezura Y, Inouye K and Takami I. 1993. Characterization of three continuous cell lines from marine fish. Journal of Aquatic Animal Health 5: 127–36.
Freshney R I. 2005. Culture of animal cells—A Manual of Basic Techniques. Pp 641. Wiley-Liss, New York USA.
Hameed A S, Parameswaran V, Shukla R, Singh I B, Thirunavukkarasu A R and Bhonde R R. 2006. Establishment and characterization of India’s first marine fish cell line from kidney of sea bass, Lates calcarifer. Aquaculture 257: 92–103.
Hamsa K M S A and Kasim H M. 1992. Growth and production potential of young grouper Epinephelus tauvina (Forskal) reared
in fixed net cages. Journal of Marine Biological Association of India 34: 271–76.
Huang X, Huang Y, Sun J, Han X and Qin Q. 2009. Characterization of two grouper Epinephelus akaara cell lines: application to studies of Singapore grouper iridovirus (SGIV) propagation and virus–host interaction. Aquaculture 292: 172–79.
Jagadis I, Ignatius B, Kandasami D and Khan A M. 2006. Embryonic and larval development of honeycomb grouper Epinephelus merra Bloch. Aquaculture Research 37: 1140–45.
Ku C C, Teng Y C, Wang C S and Lu C H. 2009. Establishment and characterization of three cell lines derived from the rockfish grouper Epinephelus quoyanus: Use for transgenic studies and cytotoxicity testing. Aquaculture 294: 147–51.
Lai Y S, John J A C, Lin C H, Guo I C, Chen S C, Fanf K, Lin C H and Chang C Y. 2003. Estabilishment of cell lines from a tropical grouper, Epinephelus awoara (Temminck & Schlegel), and their susceptibility to grouper irido- and nodaviruses. Journal of Fish Diseases 26: 31–42.
Lakra W S, Sivakumar N, Goswami M and Bhonde R R. 2006. Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch). Aquaculture Research 37: 18–24.
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.
Liu P V.1957. Survey of haemolysin production among species of Pseudomonas. Journal of Bacteriology 74: 718–27.
Luc Rougée G K, Ostrander R H and Richmond Y L. 2007. Establishment, characterization, and viral susceptibility of two cell lines derived from goldfish Carassius auratus muscle and swim bladder. Diseases of Aquatic Organisms 77: 127–35.
Nammalwar P R, Marichamy A, Regunathan and Kandasami D. 1998. Prospects of grouper culture in India. Proceedings of Workshop on National Aquaculture. pp.144–48. Aquaculture foundation of India, Chennai.
Palumbi S, Martin A, Romano S, McMillan W O, Stice L and Grabowski G. 1991. The Simple Fool’s Guide to PCR. University of Hawaii, Honolulu.
Parameswaran V, Shukla R, Bhonde R R and Sahul Hameed A S. 2006. Splenic cell line from sea bases, Lates calcarifer: Establishment and characterization. Aquaculture 261: 43–53.
Pillai V N, Murthy V S R and Mathew G. 2002. Grouper Aquaculture in India. Report of the Regional Workshop on Sustainable Seafarming and Grouper Aquaculture Pp. 224. 17– 20 April, 2000, Medan, Indonesia. Network of Aquaculture Centres in Asia Pacific, Bangkok, Thailand.
Qin Q W, Chang S F, Ngoh G H, Gibson-Kueh S, Shi C and Lam T J. 2003. Characterization of a novel ranavirus isolated from grouper Epinephelus tauvina. Diseases of Aquatic Organisms 53: 1–9.
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 Virological Methods 131: 58–64.
Ren F L, Li Y and Zhang Y. 2002. In vitro cultivation and freezing of bovine skin fibroblast cells. Journal of Yellow Cattle Science 1: 8–10.
Rimmer M A. 2004. Advances in Grouper Aquaculture. ACIAR Monograph 110. Pp. 134 (Eds) Rimmer M A, McBride and Williams K C. Australian Centre for International Agricultural Research, Canberra, Australia.
Sadovy Y J, Donaldson T J, Graham T R, McGilvray F, Muldoon G J, Phillips M J, Rimmer M A, Smith A and Yeeting B. 2003. While Stocks Last: The Live Reef Food Fish Trade. Pp.146. Asian Development Bank, Manila, Philippines.
Swaminathan T R, Lakra W S, Gopalakrishnan A, Basheer V S, Khushwaha B and Sajeela K A. 2010. Development and characterization of a new epithelial cell line PSF from caudal fin of green chromide, Etroplus suratensis. In Vitro Cellular and Developmental Biology- Animal 46 (8): 647–56.
Wang G, LaPatra S, Zeng L, Zhao Z and Lu Y. 2003. Establishment, growth, cryopreservation and species of origin identification of three cell lines from white sturgeon, Acipenser transmontanus. Methods in Cell Science 25: 211–20.
Ward R D, Zemlak T S, Innes B H, Last P R and Hebert P D N. 2005. DNA barcoding Australia’s fish species. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences 360: 1847–57.
Zhou G Z, Li Z Q, Yuan X P and Zhang Q Y. 2007. Establishment, characterization, and virus susceptibility of a new marine cell line from red spotted grouper (Epinephelus akaara). Marine Biotechnology 9: 370–76.
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2014 The Indian Journal of Animal Sciences

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.