Continuous cell culture from chick embryos inoculated with REV strain T

Oncogenic reticuloendotheliosis virus strain T (REV-T) was inoculated into 9-day-old embryonated eggs of specific pathogen free chickens. A continuous cell line was established from the liver cells of one embryo harvested the day before hatching which had the following properties: It formed loose aggregates and propagated floating in the medium. The cell doubling time was about 24 hours. Viral antigen was demonstrated in the cytoplasm by the direct immunofluorescent antibody technique. C-type particles were observed outside the cells. Virus-induced cell surface antigen was demonstrated by the immunofluorescent antibody technique. The cell line grew in soft agar. Virus production continued in the cell line for 17 months. When the cells or the culture fluid were inoculated into 1-day-old chickens, they died from neoplastic disease. The chickens which survived after infection showed abnormal feathering ("nakanuke"). The virus released from this cell-line was inoculated into 9-day-old embryonated eggs and three cell lines were established. These cell lines did not contain avian leukosis/sarcoma viruses.