Properties of an Established Cell Line from the Atlantic Croaker ( Micropogon undulatus )

Abstract

Abstract

A fibroblast-like cell line, CrF, has been developed from muscle tissue of the marine teleost fish Micropogon undulatus (the Atlantic croaker). The line was initiated by subculture of cells from explanted tissue of a single male fish. The growth medium employed contained L-15 basal medium supplemented with 15% fetal calf serum, 1 to 5% human serum, and antibiotic-antimycotic mixture. The line was characterized fully and carried through 85 subcultures. Karyological analysis showed the line to be heteroploid, with a modal chromosome number of 48 at low passage levels and 49 at high levels. All chromosomes are small and telocentric. Growth characteristics were carried out under varying conditions of incubation. Optimal growth occurred at 26 to 32° in growth medium adjusted to a NaCl molarity of 0.147 to 0.177 M (although a wider range was tolerated). A plating efficiency of up to 30% and freeze viability rates of 65 to 70% were observed. The line was shown to be susceptible to lymphocystis virus, but refractory to infectious pancreatic necrosis virus, spring viremia of carp virus, channel catfish virus, vesicular stomatitis virus, and eastern equine encephalitis virus. The identity of species of origin was confirmed by the cytotoxic antibody dye exclusion test, and the absence of microbial contamination was determined by inoculation of appropriate microbial growth media.