Abstract
The cell line has been used as a novel in vitro tool for executing several studies in life sciences. The current study aimed to develop and characterize a muscle cell culture system derived from Clarias magur. The primary muscle cell cultures derived from the caudal peduncle muscle have been successfully sub cultured up to 13 passages to establish a new muscle cell culture system known as CMM. At a temperature of 28 °C, L-15 medium supplemented with 20% FBS produced the maximum growth of muscle cells. However, muscle cells were optimized to grow at 10% FBS. To enhance the proliferation capacity of the CMM cells, a growth-promoting factor bFGF (10 ng/ml) was added, thereby reducing the time interval of passages for the subsequent cultures. DNA barcoding of the CMM cell culture system authenticated the species of origin. The cell culture system was successfully cryopreserved by a slow freezing procedure at − 80 °C with a revival efficiency of 60%.
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References
Ahmed VI, Babu VS, Chandra V, Nambi KS, Thomas J, Bhonde R, Hameed AS (2009) A new fibroblastic-like cell line from heart muscle of the Indian major carp (Catla catla): development and characterization. Aquaculture 293(3–4):180–186
Babu VS, Nambi KS, Chandra V, Ishaq Ahmed VP, Bhonde R, SahulHameed AS (2011) Establishment and characterization of a fin cell line from Indian walking catfish, Clarias batrachus (L.). J Fish Dis 34(5):355–64
Bejar J, Borrego JJ, Alvarez MC (1997) A continuous cell line from the cultured marine fish gilt-head seabream (Sparus aurata L.). Aquaculture. 150(1–2):143–53
Cooper JK, Sykes G, King S, Cottrill K, Ivanova NV, Hanner R, Ikonomi P (2007) Species identification in cell culture: a two-pronged molecular approach. In Vitro Cell Dev Biol-Anim 43:344–351
Dubey A, Goswami M, Yadav K, Sharma BS (2014) Development and characterization of a cell line WAF from freshwater shark Wallago attu. Mol Biol Rep 41:915–924
Dubey A, Goswami M, Yadav K, Mishra A, Kumar A (2015) Establishment of a novel muscle cell line from Wallago attu for in vitro study of pesticide toxicity. Gene, Cell and Tissue 2(1)
Freshney RI (2015) Culture of animal cells: a manual of basic technique and specialized applications. John Wiley & Sons
Fryer JL, Lannan CN (1994) Three decades of fish cell culture: a current listing of cell lines derived from fishes. J Tissue Cult Methods 16:87–94
Goswami M, Sharma BS, Tripathi AK, Yadav K, Bahuguna SN, Nagpure NS, Lakra WS, Jena JK (2012) Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland). Gene 500(1):140–147
Goswami M, Sharma BS, Yadav K, Bahuguna SN, Lakra WS (2014) Establishment and characterization of a piscean PCF cell line for toxicity and gene expression studies as in vitro model. Tissue Cell 46(3):206–212
Hameed AS, Parameswaran V, Shukla R, Singh IB, Thirunavukkarasu AR, Bhonde RR (2006) Establishment and characterization of India's first marine fish cell line (SISK) from the kidney of sea bass (Lates calcarifer). Aquaculture 257(1–4):92–103
Hebert PD, Ratnasingham S, De Waard JR (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc R Soc Lond Ser B: Biol Sci 270(suppl_1):S96-9
Kumar GS, Singh IB, Philip R (2001) Development of a cell culture system from the ovarian tissue of African catfish (Clarias gariepinus). Aquaculture 194(1–2):51–62
Kumar A, Singh N, Goswami M, Srivastava JK, Mishra AK, Lakra WS (2016) Establishment and characterization of a new muscle cell line of zebrafish (Danio rerio) as an in vitro model for gene expression studies. Anim Biotechnol 27(3):166–173
Lai YS, Chiou PP, Chen WJ, Chen YC, Chen CW, Chiu IS, Chen SD, Cheng YH, Chang CY (2008) Characterization of apoptosis induced by grouper iridovirus in two newly established cell lines from barramundi, Lates calcarifer (Bloch). J Fish Dis 31(11):825–834
Lakra WS, Goswami M (2011) Development and characterization of a continuous cell line PSCF from Puntius sophore. J Fish Biol 78(4):987–1001
Lakra WS, Goswami M, Rajaswaminathan T, Rathore G (2010) Development and characterization of two new cell lines from common carp, Cyprinus carpio (Linn). Biol Res 43(4):385–392
Lakra WS, Goswami M, Yadav K, Gopalakrishnan A, Patiyal RS, Singh M (2011) Development and characterization of two cell lines PDF and PDH from Puntius denisonii (Day 1865). In Vitro Cell Dev Biol-Anim 47:89–94
Mauger PE, Le Bail PY, Labbé C (2006) Cryobanking of fish somatic cells: optimizations of fin explant culture and fin cell cryopreservation. Comp Biochem Physiol b: Biochem Mol Biol 144(1):29–37
Nanda PK, Swain P, Nayak SK, Behera T, Jayasankar P, Dhama K (2014) Evaluation of different coating factors to establish cell culture from tissue explants of Indian major carp, Cirrhinus mrigala. Asian J Anim Vet Adv 9(7):395–404
Parameswaran V, Shukla R, Bhonde RR, Hameed AS (2006) Splenic cell line from sea bass, Lates calcarifer: establishment and characterization. Aquaculture 261(1):43–53
Rougée L, Ostrander GK, Richmond RH, Lu Y (2007) Establishment, characterization, and viral susceptibility of two cell lines derived from goldfish Carassius auratus muscle and swim bladder. Dis Aquat Org 77(2):127–135
Rubio N, Datar I, Stachura D, Kaplan D, Krueger K (2019) Cell-based fish: a novel approach to seafood production and an opportunity for cellular agriculture. Front Sustain Food Syst 3:43
Sambrook J (2001) Preparation and analysis of eukaryotic genomic DNA. Molecular Cloning 1:6–4
Sunil Kumar G, Bright Singh IS (2000) Development of cell culture systems from selected species of fish and prawns. Doctoral dissertation, School of Environment Studies, Faculty of Environment Studies
Wang G, Lapatra S, Zeng L, Zhao Z, Lu Y (2004) Establishment, growth, cryopreservation and species of origin identification of three cell lines from white sturgeon, Acipenser Transmontanus. Methods Cell Sci 25:211–220
Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PD (2005) DNA barcoding Australia’s fish species. Philos Trans R Soc B: Biolo Sci 360(1462):1847–1857
Yashwanth BS, Goswami M, Kooloth Valappil R, Thakuria D, Chaudhari A (2020) Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus. Sci Rep 10(1):20051
Yashwanth BS, Pinto N, Sathiyanarayanan A, Chaudhari A, Rasal KD, Goswami M (2023) Functional characterization of Labeo rohita muscle cell line for in vitro research. Mol Biol Rep 14:1–12
Zhao1 Z, Montgomery-Brock D, Lee CS, Lu Y (2004) Establishment, characterization and viral susceptibility of 3 new cell lines from snakehead, Channa striatus (Blooch). Methods Cell Sci 25:155–66
Zhao Z, Lu Y (2006) Establishment and characterization of two cell lines from bluefin trevally Caranx melampygus. Dis Aquat Org 68(2):91–100
Acknowledgements
The authors wish to express their sincere gratitude to the Director, ICAR-Central Institute of Fisheries Education, Mumbai, Maharashtra, India, and the Indian Council of Agricultural Research, New Delhi, for providing the facilities and financial support to carry out the research. The authors also acknowledge the support of the Good Food Institute, USA grant for strengthening the cell culture facility towards the "National Smart Protein Innovation Hub on Cultivated Seafood".
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This study was funded by ICAR-Central Institute of Fisheries Education, Mumbai, Maharashtra, India.
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Sekar Dhivyakumari: bench work, original draft writing, review and editing, and methodology. Aparna Chaudhari: methodology, manuscript correction and validation. Manoj P Brahmane: methodology. Dhanjit Kumar Das: methodology. Arjunan Sathiyanarayanan: assistance in cell culture. B S Yashwanth: assistance in DNA barcoding. Nevil Pinto: assistance in growth studies. Mukunda Goswami: experimental design, conceptualization, and supervision.
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All applicable institutional guidelines for the care and use of animals were followed and approved by Institutional Animal Ethics Committee (IAEC), ICAR-Central Institute of Fisheries Education, Mumbai, Maharashtra, India.
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Dhivyakumari, S., Chaudhari, A., Brahmane, M.P. et al. Development and characterization of a new muscle cell culture system from Clarias magur (Hamilton, 1822). Fish Physiol Biochem 49, 1295–1302 (2023). https://doi.org/10.1007/s10695-023-01257-7
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DOI: https://doi.org/10.1007/s10695-023-01257-7