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Establishment and characterization of an epithelial cell line from thymus of Catla catla (Hamilton, 1822)
Authors:Dharmendra K. Chaudhary  Neeraj Sood  T. Raja Swaminathan  Gaurav Rathore  P.K. Pradhan  N.K. Agarwal  J.K. Jena
Affiliation:1. National Bureau of Fish Genetic Resources, Canal Ring Road, P.O. Dilkusha, Lucknow-226 002, Uttar Pradesh, India;2. Department of Zoology, H.N.B. Garhwal University (Campus), Badshaithaul, Tehri Garhwal, Uttarakhand, India
Abstract:
A cell line, CTE, derived from catla (Catla catla) thymus has been established by explant method and subcultured for more than 70 passages over a period of 400 days. The cell line has been maintained in L-15 (Leibovitz) medium supplemented with 10% fetal bovine serum. CTE cell line consists of homogeneous population of epithelial-like cells and grows optimally at 28 °C. Karyotype analysis revealed that the modal chromosome number of CTE cells was 50. Partial amplification, sequencing and alignment of fragments of two mitochondrial genes 16S rRNA and COI confirmed that CTE cell line originated from catla. Significant green fluorescent signals were observed when the cell line was transfected with phrGFP II-N mammalian expression vector, indicating its potential utility for transgenic and genetic manipulation studies. The CTE cells showed strong positivity for cytokeratin, indicating that cell line was epithelial in nature. The flow cytometric analysis of cell line revealed a higher number of cells in S-phase at 48 h, suggesting a high growth rate. The extracellular products of Vibrio cholerae MTCC 3904 were toxic to the CTE cells. This cell line was not susceptible to fish betanodavirus, the causative agent of viral nervous necrosis in a large variety of marine fish.
Keywords:CTE cell line, catla thymus epithelial cell line   CPE, cytopathic effect   ECPs, extracellular products   MTCC, microbial type culture collection   TEC, thymic epithelial cells
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