Development, characterization and in vitro applications of a thymus cell line from Pangasianodon hypophthalmus (Sauvage 1878)
Open Access
- 9 March 2023
- journal article
- research article
- Published by Indian Council of Agricultural Research, Directorate of Knowledge Management in Agriculture in The Indian Journal of Animal Sciences
- Vol. 93 (2), 235-240
- https://doi.org/10.56093/ijans.v93i2.128796
Abstract
Pangasianodon hypophthalmus is an economically important catfish species cultured in India. In the present study, a continuous cell line from the thymus of P. hypophthalmus (PHT) was established using the explant method and subcultured 52 times since development. PHT cells showed optimal growth in L-15 medium containing 20% fetal bovine serum at 28 degrees C. The species of the cell line from striped catfish was confirmed through PCR amplification and sequencing of 16S, and COI genes. The cell line was found to be Mycoplasma free. The modal chromosome number of PHT cells was 60 (2n). Immunophenotyping using different antibodies showed the epithelial nature of the cells. Cytotoxicity of arsenic and mercury was assessed using Neutral red and MTT assay, which revealed reduced cell survival with an increase in toxicant concentration. Cells transfected successfully with the GFP reporter gene using lipofectamine reagent indicated the suitability of the cell line for expression studies. The cell line has been submitted to NRFC, Lucknow with accession no. NRFC-078 at ICAR-NBFGR, Lucknow. The developed cell line will have applications in suspected viral disease investigation, transgenic, and immunological studies.Keywords
This publication has 32 references indexed in Scilit:
- Establishment of a macrophage cell line from adherent peripheral blood mononuclear cells of Catla catlaIn Vitro Cellular & Developmental Biology – Animal, 2012
- Polymerase chain reaction-based species verification and microsatellite analysis for canine cell line validationJournal of Veterinary Diagnostic Investigation, 2011
- Development and characterization of a new epithelial cell line PSF from caudal fin of Green chromide, Etroplus suratensis (Bloch, 1790)In Vitro Cellular & Developmental Biology – Animal, 2010
- Characterization of goldfish fin cells in culture: Some evidence of an epithelial cell profileComparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2008
- Neutral red uptake assay for the estimation of cell viability/cytotoxicityNature Protocols, 2008
- DNA barcoding Australia's fish speciesPhilosophical Transactions B, 2005
- Production of recombinant protein therapeutics in cultivated mammalian cellsNature Biotechnology, 2004
- Identification and elimination of mycoplasmas in fish cell line culturesJournal of Fish Diseases, 1996
- The thymus in fish: Development and possible function in the immune responseAnnual Review of Fish Diseases, 1992
- Comparisons of two in vitro cytotoxicity assays—The neutral red (NR) and tetrazolium MTT testsToxicology in Vitro, 1988