A new fibroblast like cell line from the fry of golden mahseer Tor putitora (Ham)

Abstract

We report the development of a diploid cell line (TP-1) for the first time from golden mahseer Tor putitora, a world famous food and game fish. Thirty-day-old fry were used to prepare a primary cell culture employing trypsin digestion. The optimal conditions of nutrition and incubation for growth of the dissociated cells were determined. The [ 3H] thymidine uptake assay was employed to investigate the effect of different concentrations of fetal calf serum (FCS) and fish muscle extract (FME) on the growth of TP-1 cells and to study the cell proliferation rate at different time intervals. It was found that L-15 supplemented with 20% FCS and 10% FME at an incubation temperature of 28 °C resulted in optimal growth. The concentration of fetal calf serum was reduced to 10% after 10 subcultures. After confluency, the cells were subcultured with a split ratio of 1:2. The cells showed a fibroblast-like morphology and reached confluency on the fourth day after subculture. The cells were successfully cryopreserved and revived at passage numbers 5, 8, 15 and 18. The cells were characterized for chromosome number (2 n = 100) at 10 and 20 passages. The cell cycle analysis by FACS (Fluorescence Activated Cell Sorter) revealed that most of the cells on the first and fourth day of culture were in S-phase, indicating a high growth rate.