Development and characterization of cell line from the gill tissue of Catla catla (Hamilton, 1822) for toxicological studies

Abstract

Catla gill cell line (ICG) was established from gill tissue of Indian major carp (Catla catla), a freshwater fish cultivated in India. The cell line was maintained in Leibovitz's L-15 supplemented with 10% fetal bovine serum. These cells have been sub-cultured more than 55 passages over a period of 2years. The ICG cell line consists predominantly of epithelial-like cells. The cells were able to grow at a wide range of temperatures from 24°C to 32°C with an optimum temperature of 28°C. The growth rate of gill cells increased as the fetal bovine serum (FBS) proportion increased from 2% to 20% at 28°C with optimum growth at the concentrations of 10% or 15% FBS. Amplification of mitochondrial gene 12s rRNA using primers specific to C. catla confirmed the origin of this cell line from C. catla. The cells were successfully cryopreserved and revived at passage numbers 25, 35, 45 and 55. The cytotoxicity of three metal salts (ZnCl2, CuSO4 and CdCl2) was assessed in ICG cell line using multiple endpoints such as MTT, Neutral Red assay, Alamar Blue assay and Coomassie Blue protein assay. Acute toxicity assay on fish were conducted by exposing C. catla for 96h to three metal salts under static conditions. Statistical analysis revealed good correlation with r2=0.908-0.985 for all combinations between endpoints employed. Linear correlations between each in vitro EC50 and the in vivo LC50 data were highly significant. © 2012 Elsevier Ltd.