Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland)

Goswami, Mukunda; Sharma, B. Sharan; Tripathi, Amit Kumar; Yadav, Kamalendra; Bahuguna, S. N.; Nagpure, N. S.; Lakra, W. S.; Jena, J. K.

doi:10.1016/j.gene.2012.03.016

Cited by 17 publications

(10 citation statements)

References 49 publications

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“…To overcome the problem, the culture media L-15 was supplemented with different concentrations of 0.2 M sodium chloride solution, and it was found that OCF cells grew well and formed a complete monolayer at 2 mM NaCl. The fish cell line can grow in a wide range of temperatures from 24 to 32 °C 34 , 41 – 43 . In the present study, the most suitable temperature for optimum growth and proliferation of the developed OCF cell line was revealed to be 28 °C which showed conformity with other marine fish cell cultures 44 .…”

Section: Discussionmentioning

confidence: 99%

“…The high proliferation of cells and plating efficiency observed in the cell lines is expressive of a transformed characteristic or genotypic change 46 . Goswami et al reported 64% of plating efficiency seeded at 1000 cells per flask 43 . The gene sequence of both COI and 16S rRNA obtained from the OCF cells revealed the similarity of 99–100% with the known sequences of A. ocellaris submitted in NCBI, Genbank Database.…”

Section: Discussionmentioning

confidence: 99%

“…The cells were incubated at optimum pH 7.4 and temperature 28 °C. The cultures were monitored daily and subcultured upon reaching 80% confluency 34,43 .…”

Section: Subculture and Maintenancementioning

confidence: 99%

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Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus

Yashwanth

Goswami

Valappil

et al. 2020

Sci Rep

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Amphiprion ocellaris (ocellaris clownfish) is one of the most commercially important marine ornamental fish. A cell line designated as OCF was developed for the first time from the caudal fin of this fish species. The cell line was maintained in Leibovitz’s—15 medium supplemented with 15% FBS (Fetal Bovine Serum) and was successfully subcultured up to 34 passages. The cell line was authenticated by sequencing mitochondrial cytochrome C oxidase subunit I (COI) and 16S rRNA genes. The growth rate of the OCF cell line was maximum in medium containing 20% FBS and 1% of 0.2 M NaCl at 28 °C. Chromosome analysis revealed 48 diploid chromosomes. The OCF cell line was transfected with the pMaxGFP plasmid vector with 7% efficiency and GFP expression was observed. The OCF cell line was used for testing nervous necrosis virus (NNV) susceptibility. Cytopathic effect (CPE) was observed in terms of plaque formation after virus inoculation. Nested PCR confirmed the susceptibility of the OCF cell line to NNV. The cell line was successfully cryopreserved by a slow freezing procedure at − 80 °C with a revival efficiency of 70–75%. The study revealed that the OCF cell line would be useful for virological studies. In addition, the cell line would play an important role as an in vitro tool for carrying out toxicological and biotechnological studies.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus

Yashwanth

Goswami

Valappil

et al. 2020

Sci Rep

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“…Fish-cell lines have proven to be powerful tools in fish biological and pathological research, since the establishment of the RTG-2 continuous gonad cell line from O. mykiss (Wolf & Quimby, 1962). They can provide important in vitro systems for investigations in fish immunology (Clem et al, 1996;Goswami et al, 2012), toxicology (Papis et al, 2011;Altay et al, 2012), endocrinology (Aluru & Vijayan, 2009;Thomas, 2012), virology (Ruiz et al, 2009;Kim et al, 2012), functional genomics (Laing & Hansen, 2011), physiology (Tafalla et al, 2006), cytophysiology (Maclean, 2011), genetic regulation (Castro et al, 2010;Holopainen et al, 2012) and carcinogenesis (Salinas et al, 2008). To date, however, most fish hepatocyte lines have been used for fish toxicology and immunology study, very few for research on lipid metabolism.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Liu

Zhang

Dong

et al. 2017

Journal of Fish Biology

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A hepatocyte line was established from the liver of white-spotted spinefoot Siganus canaliculatus to study the biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFA). The cells from the line, designated S. canaliculatus hepatocyte line (SCHL), grew and multiplied well in Dulbecco's modified Eagle's medium (DMEM)-F12 medium supplemented with 20 mM 4-(2-hydroxyethyl) piperazine-1-ethanesulphonic acid (HEPES), 10% foetal bovine serum (FBS) and 0·5% rainbow trout Oncorhychus mykiss serum at 28° C, showing an epithelial-like morphology and the normal chromosome number of 48 (2n) and have been subcultured for over 60 passages. The identity of the hepatocytes was confirmed by periodic acid Schiff (PAS) staining. The mRNA expression of all genes encoding the key enzymes for LC-PUFA biosynthesis including two desaturases (Δ4 Fad and Δ6-Δ5 Fad) and two elongases (Elovl4 and Elovl5), were detected in all cells from passages 5 to 60 and their expression levels became stable after passage 35 and showed responses to various PUFA incubation. This is similar to the situation determined in the liver of S. canaliculatus that were fed diets containing different fatty acids. These results indicated that SCHL was successfully established and can provide an in vitro tool to investigate lipid metabolism and regulatory mechanisms of LC-PUFA biosynthesis in teleosts, especially marine species.

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“…The most important cultivable coldwater species include Neolissoceillis hexagonolepis, Puntius (Tor) chelynoides, T. putitora, T. tor among the mahseers, Schizothorax richardsonii, Schizothorax plagiostomous and Schizothoraichthys progastus among the snowtrout (Singh et al, 1993). Only a few cell lines have been developed from warm water fish species such as catfish and from coldwater fish species such as snow trouts and mahseers , Goswami et al, 2012. Although there are about 11 reports on cell cultures from indigenous fish of India, information on their utilization and availability is lacking when in vitro studies related to fish virology and transgenesis require cell lines from Indian fish .…”

Section: Introductionmentioning

confidence: 99%

Development of primary cell culture from the caudal fin region of snow trout Schizothorax plagiostomus (Heckel)

Chowdhary

Bahuguna

2014

ECJ

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A primary cell culture has been established from the Snow trout Schizothorax plagiostomus. Primary cultures have developed from caudal fin tissues by explant culture technique. Cell outgrowth has been obtained from the caudal fin explant after 5-7 days of explant culture. The culture medium used for the experiment was Leibovitz-15 supplemented with 20% Fetal Bovine Serum. Radiation of cells from the explants started after 2 to 3 days. Both fibroblast like and epithelial like cells were noticed but fibroblast like cells dominated as the growth progressed.

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Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland)

Cited by 17 publications

References 49 publications

Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus

Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Development of primary cell culture from the caudal fin region of snow trout Schizothorax plagiostomus (Heckel)

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