Abstract
A cell line (SICH) was established from heart muscle of Indian major carp (Catla catla), a freshwater fish cultivated in India. The cell line was maintained in Leibovitz's L-15 supplemented with 15% fetal bovine serum. These cells have been sub-cultured more than 130 times over a period of two years. The SICH cell line consists predominantly of fibroblastic-like cells. The cells were able to grow at a wide range of temperatures from 24 degrees C to 37 degrees C with an optimum temperature of 28 degrees C. The growth rate of heart cells increased as the FBS proportion increased from 2 to 20% at 28 degrees C with optimum growth at the concentrations of 10 or 15% FBS. PCR amplification of mitochondrial 12s rRNA using primers specific to C. catla confirmed the origin of this cell line from catla. The cells were successfully cryopreserved and revived at passage numbers 20, 35, 70 and 120. The cell cycle analysis by FACS (Fluorescence Activated Cell Sorter) revealed that most of the cells on the second day of culture were in S-phase, indicating a high growth rate. When the SICH cells were transfected with pEGFP using lipofectamine 2000 or chitosan nanoparticle, fluorescent signals were observed, suggesting that the SICH cell line could be a useful tool for transgenic and genetic manipulation studies. The bacterial extracellular products from Aeromonas sp., or Vibrio anguillarum were found to be toxic to this cell line. This cell line was further characterized by immunocytochemistry using confocal laser scanning microscopy. In addition, an attempt was also made to use this cell line for genotoxicity studies. (C) 2009 Elsevier B.V. All rights reserved.