Establishment and characterization of 13 cell lines from a green turtle (Chelonia mydas) with fibropapillomas

Thirteen cell lines were established and characterized from brain, kidney, lung, spleen, heart, liver. gall bladder urinary bladder, pancreas, testis, skin, and periorbital and tumor tissues of an immature male green turtle ( Chelonia mydas) with fibropapillomas. Cell lines were optimally maintained at 30 degrees C in RPMI 1640 medium supplemented with 10% fetal bovine seru m. Propagation of the turtle cell lines was serum dependent, and plating ef ficiencies ranged from 13 to 37%. The cell lines, which have been subcultiv ated more than 20 times, had a doubling time of approximately 30 to 36 h. W hen tested for their sensitivity to several fish viruses, most of the cell lines were susceptible to a rhabdovirus, spring viremia carp virus. but ref ractory; to channel catfish virus (a herpesvirus), infectious pancreatic ne crosis virus (a birnavirus), and two other fish rhabdoviruses, infectious h ematopoietic necrosis virus and viral hemorrhagic septicemia virus. During in vitro subcultivation: tumor-like cell aggregates appeared in cell lines derived from lungs, testis, and periorbital and tumor tissues, and small, n aked intranuclear virus particles were detected by thin-section electron mi croscopy. These cell lines are currently being used in attempts to isolate the putative etiologic virus of green turtle fibropapilloma.