Development and characterization of a fibroblastic-like cell line from caudal fin of the red-line torpedo, Puntius denisonii (Day) (Teleostei: Cyprinidae)

摘要

A fibroblastic-like cell line was established from the ornamental fish, red-line torpedo (Puntius denisonii). The red-line torpedo fin (RTF) cell line is being maintained in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum (FBS) for over 1 year at 28?°C on a continuous basis in normal atmosphere. The growth rate of RTF cells increased as the FBS proportion increased from 5% to 20% at 28?°C with optimum growth at the concentrations of 10% FBS. The morphology of RTF cell was predominantly fibroblastic like. Propagation of these cell lines was serum dependent, with a low plating efficiency (<15%). Karyotyping analysis of RTF cells at the 25th passage indicated that the modal chromosome number was 2n=50. The cell line was cryopreserved in liquid nitrogen at ?196?°C and could be recovered from storage after 6 months with good cell viability. Polymerase chain reaction amplification of a fragment of two mitochondrial genes, 16S rRNA and CO1, confirmed the identity of these cell lines with those reported from this animal species, confirming that the cell lines originated from P. denisonii. The bacterial extracellular products from Vibrio cholerae MTCC3904 and Aeromonas hydrophila were found to be toxic to RTF. The cell lines were not susceptible to viral nervous necrosis virus, a marine fish virus.